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Research Paper
Cell culture of taxus as a source of the antineoplastic drug taxol and related taxanes.
- Arthur G. Fett-Neto
- Frank DiCosmo
Biological Oxidation of Hydrochlorofluorocarbons (HCFCs) by a Methanotrophic Bacterium
- Mary F. DeFlaun
- Burt D. Ensley
- Robert J. Steffan
Solubilization and Activity of Proteins in Compressible-Fluid Based Microemulsions
- Guadalupe Ayala
- Sanjay V. Kamat
- Alan J. Russell
An Algorithmically Optimized Combinatorial Library Screened by Digital Imaging Spectroscopy
- Ellen R. Goldman
- Douglas C. Youvan
Hyperthermostable Variants of a Highly Thermostable Alpha-Amylase
- Philippe Joyet
- Nathalie Declerck
- Claude Gaillardin
Fertile, Transgenic Oat Plants
- David A. Somers
- Howard W. Rines
- William R. Bushnell
Comparison of Coat Protein-Mediated and Genetically-Derived Resistance in Cucumbers to Infection by Cucumber Mosaic Virus Under Field Conditions with Natural Challenge Inoculations by Vectors
- Dennis Gonsalves
- Jerry L. Slightom
Controlled Antibody Delivery Systems
- Jill K. Sherwood
- Richard B. Dause
- W. Mark Saltzman
Rescuing Transgene Expression by Co-Integration
- A. J. Clark
- J. P. Simons
Trypanosoma Cruzi Flagellar Repetitive Antigen Expression by Recombinant Baculovirus: Towards an Improved Diagnostics Reagent for Chagas' Disease
- Claudia N. Duarte dos Santos
- Marco A. Krieger
- Ricardo Galler
“Primatization” of Recombinant Antibodies for Immunotherapy of Human Diseases: A Macaque/Human Chimeric Antibody Against Human CD4
- Roland Newman
- James Alberts
- Nabil Hanna
The Two Major Xylanases from Trichoderma Reesei : Characterization of Both Enzymes and Genes
- Anneli Törrönen
- Robert L. Mach
- Christian P. Kubicek
Virus Resistant Papaya Plants Derived from Tissues Bombarded with the Coat Protein Gene of Papaya Ringspot Virus
- Maureen M. M. Fitch
- Richard M. Manshardt
- John C. Sanford
Effect of Glycosylation on Properties of Soluble Interferon Gamma Receptors Produced in Prokaryotic and Eukaryotic Experession Systems
- Michael Fountoulakis
- Reiner Gentz
High Level Expression of Streptokinase in Escherichia Coli
- M. P. Estrada
- L. Hernandez
Baculovirus Expression of Alkaline Phosphatase as a Reporter Gene for Evaluation of Production, Glycosylation and Secretion
- T. R. Davis
- K. Munkenbeck Trotter
Characterization of RNA–Mediated Resistance to Tomato Spotted Wilt Virus in Transgenic Tobacco Plants
- Peter de Haan
- Jan J. L. Gielen
- Rob Goldbach
Non–Neutralizing Monoclonal Antibodies Against RAS GTPase–Activating Protein: Production, Characterization and Use in an Enzyme Immunometric Assay
- G. Y. Zhang
- M. N. Thang
Construction, Bacterial Expression and Characterization of a Bifunctional Single–Chain Antibody–Phosphatase Fusion Protein Targeted to the Human ERBB–2 Receptor
- Winfried Wels
- Ina-Maria Harwerth
- Nancy E. Hynes
High Level Expression of a Chimeric Anti–Ganglioside GD2 Antibody: Genomic Kappa Sequences Improve Expression in COS and CHO Cells
- Lynette A. Fouser
- Stephen L. Swanberg
- Gerard E. Riedel
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Page 1 of 34
A fully human connective tissue growth factor blocking monoclonal antibody ameliorates experimental rheumatoid arthritis through inhibiting angiogenesis
Connective tissue growth factor (CTGF) plays a pivotal role in the pathogenesis of rheumatoid arthritis (RA) by facilitating angiogenesis and is a promising therapeutic target for RA treatment. Herein, we gene...
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Black glucose-releasing silicon elastomer rings for fed-batch operation allow measurement of the oxygen transfer rate from the top and optical signals from the bottom for each well of a microtiter plate
In industrial microbial biotechnology, fed-batch processes are frequently used to avoid undesirable biological phenomena, such as substrate inhibition or overflow metabolism. For targeted process development, ...
In vitro culture of bovine fibroblasts using select serum-free media supplemented with Chlorella vulgaris extract
Standard cell culture practices require the addition of animal-derived serum to culture media to achieve adequate cell growth. Typically, 5–10% by volume of fetal bovine serum (FBS) is used, which accounts for...
Biochemical characterization and insights into the potency of the acidic Aspergillus niger NRC114 purified α-galactosidase in removing raffinose family oligosaccharides from soymilk yogurt
Because humans lack α-galactosidase, foods containing certain oligosaccharides from the raffinose family, such as soybeans and other legumes, may disrupt digestion and cause flatulence.
Production of rhamnolipid biosurfactants in solid-state fermentation: process optimization and characterization studies
Rhamnolipids are a group of the extracellular microbial surface-active molecules produced by certain Pseudomonas species with various environmental and industrial applications. The goal of the present research wa...
Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts
Chloroplast transformation is a robust technology for the expression of recombinant proteins. Various types of pharmaceutical proteins including growth factors have been reported in chloroplasts via chloroplas...
Enhancing docosahexaenoic acid production of Schizochytrium sp. by optimizing fermentation using central composite design
Docosahexaenoic acid (DHA) can improve human and animal health, particularly including anti-inflammatory, antioxidant, anticancer, neurological, and visual functions. Schizochytrium sp. is a marine heterotrophic ...
Rescue of an enterotropic Newcastle disease virus strain ZM10 from cloned cDNA and stable expressing an inserted foreign gene
Newcastle disease virus (NDV) strain ZM10, a typical enterotropic avirulent vaccine strain, has been widely used in China for chickens against Newcastle disease. To elucidate its enterotropic mechanism and dev...
Correction: Identifcation, expression, and purifcation of DNA cytosine 5-methyltransferases with short recognition sequences
The original article was published in BMC Biotechnology 2022 22 :33
Development of a microbial protease for composting swine carcasses, optimization of its production and elucidation of its catalytic hydrolysis mechanism
Dead swine carcass composting is an excellent method for harmless treatment and resource utilization of swine carcass. However, poor biodegradation ability of traditional composting results in poor harmless tr...
Toxic effects of trace phenol/guanidine isothiocyanate (P/GI) on cells cultured nearby in covered 96-well plates
A mixture of phenol and guanidine isothiocyanate (“P/GI”, the principal components of TRIzol™ and similar products) is routinely used to isolate RNA, DNA, and proteins from a single specimen. In time-course ex...
Modeling and optimizing in vitro percentage and speed callus induction of carrot via Multilayer Perceptron-Single point discrete GA and radial basis function
Callus induction is the first step in optimizing plant regeneration. Fit embryogenesis and shooting rely on callus induction. In addition, using artificial intelligence models in combination with an algorithm ...
Identification, expression, and purification of DNA cytosine 5-methyltransferases with short recognition sequences
DNA methyltransferases (MTases) are enzymes that induce methylation, one of the representative epigenetic modifications of DNA, and are also useful tools for analyzing epigenomes. However, regarding DNA cytosi...
The Correction to this article has been published in BMC Biotechnology 2022 22 :37
Preparation of a novel monoclonal antibody against active components of PHA-L from Phaseolus vulgaris and its functional characteristics
Leukocyte phytohemagglutinin (PHA-L), derived from the L4 tetramer of PHA, has been frequently employed as a mitogen to induce T lymphocyte proliferation in vitro. The biological application of PHA-L in cancer...
Identification of a novel fully human anti-toxic shock syndrome toxin (TSST)-1 single-chain variable fragment antibody averting TSST-1-induced mitogenesis and cytokine secretion
Staphylococcal superantigens are virulence factors that help the pathogen escape the immune system and develop an infection. Toxic shock syndrome toxin (TSST)-1 is one of the most studied superantigens whose r...
Gene cloning and molecular characterization of a thermostable chitosanase from Bacillus cereus TY24
An important conceptual advance in health and the environment has been recognized that enzymes play a key role in the green processing industries. Of particular interest, chitosanase is beneficial for recyclin...
Investigation of neomycin biodegradation conditions using ericoid mycorrhizal and white rot fungal species
In the search for methods to biodegrade recalcitrant compounds, the use of saprotrophic fungi and white rot fungi, in particular belonging to the phylum Basidiomycota, has gained interest. This group of fungi ...
Automatic identification of benign pigmented skin lesions from clinical images using deep convolutional neural network
We aimed to develop a computer-aided detection (CAD) system for accurate identification of benign pigmented skin lesions (PSLs) from images captured using a digital camera or a smart phone.
A new method for screening acute/chronic lymphocytic leukemia: dual-label time-resolved fluorescence immunoassay
Lymphocytic leukemia (LL) is a primary malignant tumor of hematopoietic tissue, which seriously affects the health of children and the elderly. The study aims to establish a new detection method for screening ...
Production of surfactant-stable keratinase from Bacillus cereus YQ15 and its application as detergent additive
With the growing concern for the environment, there are trends that bio-utilization of keratinous waste by keratinases could ease the heavy burden of keratinous waste from the poultry processing and leather in...
Efficient gene delivery into the embryonic chicken brain using neuron-specific promoters and in ovo electroporation
The chicken in ovo model is an attractive system to explore underlying mechanisms of neural and brain development, and it is important to develop effective genetic modification techniques that permit analyses ...
Metabolic engineering of energycane to hyperaccumulate lipids in vegetative biomass
The metabolic engineering of high-biomass crops for lipid production in their vegetative biomass has recently been proposed as a strategy to elevate energy density and lipid yields for biodiesel production. En...
940 nm diode laser induced differentiation of human adipose derived stem cells to temporomandibular joint disc cells
Temporomandibular disorder (TMD) refers to a group of disorders that affect temporomandibular joint (TMJ) and its associated muscles with very limited treatment options. Stem cell research is emerging as one o...
Development of a human phage display-derived anti-PD-1 scFv antibody: an attractive tool for immune checkpoint therapy
The PD-1 checkpoint pathway plays a major role in tumor immune evasion and the development of the tumor microenvironment. Clinical studies show that therapeutic antibodies blocking the PD-1 pathway can restore...
Generation of a recombinant antibody for sensitive detection of Pseudomonas aeruginosa
Pseudomonas aeruginosa ( P. aeruginosa ) is a major pathogen that causes nosocomial infections and often exhibits antibiotic resistance. Therefore, the development of an accurate method for detecting P. aeruginosa ...
Oleic acid based experimental evolution of Bacillus megaterium yielding an enhanced P450 BM3 variant
Unlike most other P450 cytochrome monooxygenases, CYP102A1 from Bacillus megaterium (BM3) is both soluble and fused to its redox partner forming a single polypeptide chain. Like other monooxygenases, it can catal...
Generation of genome-edited dogs by somatic cell nuclear transfer
Canine cloning technology based on somatic cell nuclear transfer (SCNT) combined with genome-editing tools such as CRISPR-Cas9 can be used to correct pathogenic mutations in purebred dogs or to generate animal...
rocF affects the production of tetramethylpyrazine in fermented soybeans with Bacillus subtilis BJ3-2
Tetramethylpyrazine (TTMP) is a flavoring additive that significantly contributes to the formation of flavor compounds in soybean-based fermented foods. Over recent years, the application of TTMP in the food i...
Development of an efficient veterinary rabies vaccine production process in the avian suspension cell line AGE1.CR.pIX
Mass vaccination of dogs as important rabies reservoir is proposed to most effectively reduce and eliminate rabies also in humans. However, a minimum coverage of 70% needs to be achieved for control of the dis...
Oil–water partition coefficient preparation and detection in the dihydroartemisinin self-emulsifying drug delivery system
The aim of the present study is to increase the solubility of dihydroartemisinin (DHA) using the self-emulsifying drug delivery system (SEDDS).
αvβ3-targeted sEVs for efficient intracellular delivery of proteins using MFG-E8
Small extracellular vesicles (sEVs) are nanometer-sized membranous particles shed by many types of cells and can transfer a multitude of cargos between cells. Recent studies of sEVs have been focusing on their...
Introduction of loxP sites by electroporation in the mouse genome; a simple approach for conditional allele generation in complex targeting loci
The discovery of the CRISPR-Cas9 system and its applicability in mammalian embryos has revolutionized the way we generate genetically engineered animal models. To date, models harbouring conditional alleles (i...
Method for quantification of porcine type I interferon activity using luminescence, by direct and indirect means
Type I interferons are widely used in research applications and as biotherapeutics. Current assays used to measure interferon concentrations, such as plaque reduction assays and ELISA, are expensive, technical...
A copper switch for inducing CRISPR/Cas9-based transcriptional activation tightly regulates gene expression in Nicotiana benthamiana
CRISPR-based programmable transcriptional activators (PTAs) are used in plants for rewiring gene networks. Better tuning of their activity in a time and dose-dependent manner should allow precise control of ge...
Isolation and identification of a feather degrading Bacillus tropicus strain Gxun-17 from marine environment and its enzyme characteristics
Feathers are the most abundant agricultural waste produced by poultry farms. The accumulation of a large number of feathers not only seriously pollutes the environment but also causes the waste of protein reso...
A simplified Gibson assembly method for site directed mutagenesis by re-use of standard, and entirely complementary, mutagenesis primers
Site-directed mutagenesis (SDM) is a key method in molecular biology; allowing to modify DNA sequences at single base pair resolution. Although many SDM methods have been developed, methods that increase effic...
Efficient extraction and antioxidant activity of polyphenols from Antrodia cinnamomea
Antrodia cinnamomea , a rare medicinal fungus, has been increasingly studied in recent years because of its abundant secondary metabolites which are beneficial to humans. However, there is a lack of research on it...
Development of a CHO cell line for stable production of recombinant antibodies against human MMP9
Human matrix metalloproteinase 9 (hMMP9) is a biomarker in several diseases, including cancer, and the need for developing detectors and inhibitors of hMMP9 is increasing. As an antibody against hMMP9 can be s...
An efficient and specific CRISPR-Cas9 genome editing system targeting soybean phytoene desaturase genes
Genome editing by CRISPR/Cas9 has become a popular approach to induce targeted mutations for crop trait improvement. Soybean ( Glycine max L. Merr.) is an economically important crop worldwide. Although gene editi...
One-step fermentation for producing xylo-oligosaccharides from wheat bran by recombinant Escherichia coli containing an alkaline xylanase
One-step fermentation is a cheap way to produce xylo-oligosaccharides (XOS), where production of xylanases and XOS is integrated into a single process. In spite of cost advantage, one-step fermentation is stil...
Self-limiting fall armyworm: a new approach in development for sustainable crop protection and resistance management
The fall armyworm, Spodoptera frugiperda , is a significant and widespread pest of maize, sorghum, rice, and other economically important crops. Successful management of this caterpillar pest has historically reli...
Biological modification of pentosans in wheat B starch wastewater and preparation of a composite film
Petrochemical resources are becoming increasingly scarce, and petroleum-based plastic materials adversely impact the environment. Thus, replacement of petroleum-based materials with new and effective renewable...

A study on the processing technology for Rhizoma Coptidis
The present study intends to optimize the processing technology for the wine-processing of Rhizoma Coptidis, using alkaloids as indicators.
Tenebrio molitor : possible source of polystyrene-degrading bacteria
The excessive use of polystyrene as a packaging material has resulted in a rise in environmental pollution. Polystyrene waste has continually increased water pollution, soil pollution and the closing of landfi...
Media optimization for SHuffle T7 Escherichia coli expressing SUMO-Lispro proinsulin by response surface methodology
SHuffle is a suitable Escherichia coli ( E. coli ) strain for high yield cytoplasmic soluble expression of disulfide-bonded proteins such as Insulin due to its oxidative cytoplasmic condition and the ability to cor...
Expression, purification and characterisation of a human anti-CDK4 single-chain variable fragment antibody
Cyclin-dependent kinase 4 (CDK4) when hyperactivated drives development and maintenance of most tumour types, thus prompting its use as an essential cancer treatment target and a diagnostic tool. Target-bindin...
Enzymatic degradation is an effective means to reduce aflatoxin contamination in maize
Aflatoxins are carcinogenic compounds produced by certain species of Aspergillus fungi. The consumption of crops contaminated with this toxin cause serious detrimental health effects, including death, in both liv...
Chloroplast genome sequencing based on genome skimming for identification of Eriobotryae Folium
Whole chloroplast genome (cpDNA) sequence is becoming widely used in the phylogenetic studies of plant and species identification, but in most cases the cpDNA were acquired from silica gel dried fresh leaves. ...
Antibacterial effect of cerium oxide nanoparticle against Pseudomonas aeruginosa
Antibiotics have been widely used for the treatment of bacterial infections for decades. However, the rapid emergence of antibiotic-resistant bacteria has created many problems with a heavy burden for the medi...
Computational modeling of PET tracer distribution in solid tumors integrating microvasculature
We present computational modeling of positron emission tomography radiotracer uptake with consideration of blood flow and interstitial fluid flow, performing spatiotemporally-coupled modeling of uptake and int...
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Journal of Biotechnology
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Most Downloaded Articles
The most downloaded articles from Journal of Biotechnology in the last 90 days.
Potential of microalgae as a sustainable feed ingredient for aquaculture
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Enhanced CRISPR/Cas9-mediated precise genome editing by improved design and delivery of gRNA, Cas9 nuclease, and donor DNA
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Bacterial enzymes involved in lignin degradation
Gonzalo de Gonzalo, Dana I. Colpa, Mohamed H.M. Habib, Marco W. Fraaije Open Access October 2016
Polyhydroxyalkanoate bio-production and its rise as biomaterial of the future
Tania Palmeiro-Sánchez, Vincent O’Flaherty, Piet N.L. Lens Open Access March 2022
Impacts of fast production of afucosylated antibodies and Fc mutants in ExpiCHO-S™ for enhancing FcγRIIIa binding and NK cell activation
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In vitro models of medulloblastoma: Choosing the right tool for the job
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Astrid Weninger, Anna-Maria Hatzl and 3 more Open Access October 2016
Therapeutic glycoprotein production in mammalian cells
Marie-Eve Lalonde, Yves Durocher Open Access June 2017
The use of bacteriophages to biocontrol oral biofilms
Szymon P. Szafrański, Andreas Winkel, Meike Stiesch Open Access May 2017
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Djordje Fira, Ivica Dimkić and 3 more November 2018
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Recombinant Protein L: Production, Purification and Characterization of a Universal Binding Ligand
Stefan Kittler, Julian Ebner and 9 more Open Access November 2022
lac operon induction in Escherichia coli: Systematic comparison of IPTG and TMG induction and influence of the transacetylase LacA
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Small-scale GMP production of plasmid DNA using a simplified and fully disposable production method
Noor A.M. Bakker, Renate de Boer and 6 more Open Access December 2019
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Emily M. Anderson, Amanda Haupt and 9 more Open Access October 2015
Extraction of astaxanthin from microalga Haematococcus pluvialis in red phase by using generally recognized as safe solvents and accelerated extraction
Antonio Molino, Juri Rimauro and 9 more Open Access October 2018
Recent advances in CRISPR/Cas9-mediated knock-ins in mammalian cells
Mehdi Banan January 2020
Advanced genetic strategies for recombinant protein expression in Escherichia coli
Hans Peter Sørensen, Kim Kusk Mortensen January 2005
Advanced PHB fermentation strategies with CO2-derived organic acids
Elodie Vlaeminck, Koen Quataert and 3 more Open Access January 2022
Flashing lights affect the photophysiology and expression of carotenoid and lipid synthesis genes in Nannochloropsis gaditana
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Environmental analysis of plastic production processes: Comparing petroleum-based polypropylene and polyethylene with biologically-based poly-β-hydroxybutyric acid using life cycle analysis
K.G. Harding, J.S. Dennis, H. von Blottnitz, S.T.L. Harrison May 2007
Bioinformatics for NGS-based metagenomics and the application to biogas research
Sebastian Jünemann, Nils Kleinbölting and 10 more Open Access November 2017
Biotechnology Research Paper Topics

This collection of biotechnology research paper topics provides the list of 10 potential topics for research papers and overviews the history of biotechnology.

1. Animal Breeding: Genetic Methods
Modern animal breeding relies on scientific methods to control production of domesticated animals, both livestock and pets, which exhibit desired physical and behavioral traits. Genetic technology aids animal breeders to attain nutritional, medical, recreational, and fashion standards demanded by consumers for animal products including meat, milk, eggs, leather, wool, and pharmaceuticals. Animals are also genetically designed to meet labor and sporting requirements for speed and endurance, conformation and beauty ideals to win show competitions, and intelligence levels to perform obediently at tasks such as herding, hunting, and tracking. By the late twentieth century, genetics and mathematical models were appropriated to identify the potential of immature animals. DNA markers indicate how young animals will mature, saving breeders money by not investing in animals lacking genetic promise. Scientists also successfully transplanted sperm-producing stem cells with the goal of restoring fertility to barren breeding animals. At the National Animal Disease Center in Ames, Iowa, researchers created a gene-based test, which uses a cloned gene of the organism that causes Johne’s disease in cattle in order to detect that disease to avert epidemics. Researchers also began mapping the dog genome and developing molecular techniques to evaluate canine chromosomes in the Quantitative Trait Loci (QTL). Bioinformatics incorporates computers to analyze genetic material. Some tests were developed to diagnose many of several hundred genetic canine diseases including hip dysplasia and progressive retinal atrophy (PRA). A few breed organizations modified standards to discourage breeding of genetically flawed animals and promote heterozygosity.
2. Antibacterial Chemotherapy
In the early years of the twentieth century, the search for agents that would be effective against internal infections proceeded along two main routes. The first was a search for naturally occurring substances that were effective against microorganisms (antibiosis). The second was a search for chemicals that would have the same effect (chemotherapy). Despite the success of penicillin in the 1940s, the major early advances in the treatment of infection occurred not through antibiosis but through chemotherapy. The principle behind chemotherapy was that there was a relationship between chemical structure and pharmacological action. The founder of this concept was Paul Erhlich (1854–1915). An early success came in 1905 when atoxyl (an organic arsenic compound) was shown to destroy trypanosomes, the microbes that caused sleeping sickness. Unfortunately, atoxyl also damaged the optic nerve. Subsequently, Erhlich and his co-workers synthesized and tested hundreds of related arsenic compounds. Ehrlich was a co-recipient (with Ilya Ilyich Mechnikov) of the Nobel Prize in medicine in 1908 for his work on immunity. Success in discovering a range of effective antibacterial drugs had three important consequences: it brought a range of important diseases under control for the first time; it provided a tremendous stimulus to research workers and opened up new avenues of research; and in the resulting commercial optimism, it led to heavy postwar investment in the pharmaceutical industry. The therapeutic revolution had begun.
3. Artificial Insemination and in Vitro Fertilization
Artificial insemination (AI) involves the extraction and collection of semen together with techniques for depositing semen in the uterus in order to achieve successful fertilization and pregnancy. Throughout the twentieth century, the approach has offered animal breeders the advantage of being able to utilize the best available breeding stock and at the correct time within the female reproductive cycle, but without the limitations of having the animals in the same location. AI has been applied most intensively within the dairy and beef cattle industries and to a lesser extent horse breeding and numerous other domesticated species.
Many of the techniques involved in artificial insemination would lay the foundation for in vitro fertilization (IVF) in the latter half of the twentieth century. IVF refers to the group of technologies that allow fertilization to take place outside the body involving the retrieval of ova or eggs from the female and sperm from the male, which are then combined in artificial, or ‘‘test tube,’’ conditions leading to fertilization. The fertilized eggs then continue to develop for several days ‘‘in culture’’ until being transferred to the female recipient to continue developing within the uterus.
4. Biopolymers
Biopolymers are natural polymers, long-chained molecules (macromolecules) consisting mostly of a repeated composition of building blocks or monomers that are formed and utilized by living organisms. Each group of biopolymers is composed of different building blocks, for example chains of sugar molecules form starch (a polysaccharide), chains of amino acids form proteins and peptides, and chains of nucleic acid form DNA and RNA (polynucleotides). Biopolymers can form gels, fibers, coatings, and films depending on the specific polymer, and serve a variety of critical functions for cells and organisms. Proteins including collagens, keratins, silks, tubulins, and actin usually form structural composites or scaffolding, or protective materials in biological systems (e.g., spider silk). Polysaccharides function in molecular recognition at cell membrane surfaces, form capsular barrier layers around cells, act as emulsifiers and adhesives, and serve as skeletal or architectural materials in plants. In many cases these polymers occur in combination with proteins to form novel composite structures such as invertebrate exoskeletons or microbial cell walls, or with lignin in the case of plant cell walls.
The use of the word ‘‘cloning’’ is fraught with confusion and inconsistency, and it is important at the outset of this discussion to offer definitional clarification. For instance, in the 1997 article by Ian Wilmut and colleagues announcing the birth of the first cloned adult vertebrate (a ewe, Dolly the sheep) from somatic cell nuclear transfer, the word clone or cloning was never used, and yet the announcement raised considerable disquiet about the prospect of cloned human beings. In a desire to avoid potentially negative forms of language, many prefer to substitute ‘‘cell expansion techniques’’ or ‘‘therapeutic cloning’’ for cloning. Cloning has been known for centuries as a horticultural propagation method: for example, plants multiplied by grafting, budding, or cuttings do not differ genetically from the original plant. The term clone entered more common usage as a result of a speech in 1963 by J.B.S. Haldane based on his paper, ‘‘Biological possibilities for the human species of the next ten-thousand years.’’ Notwithstanding these notes of caution, we can refer to a number of processes as cloning. At the close of the twentieth century, such techniques had not yet progressed to the ability to bring a cloned human to full development; however, the ability to clone cells from an adult human has potential to treat diseases. International policymaking in the late 1990s sought to distinguish between the different end uses for somatic cell nuclear transfer resulting in the widespread adoption of the distinction between ‘‘reproductive’’ and ‘‘therapeutic’’ cloning. The function of the distinction has been to permit the use (in some countries) of the technique to generate potentially beneficial therapeutic applications from embryonic stem cell technology whilst prohibiting its use in human reproduction. In therapeutic applications, nuclear transfer from a patient’s cells into an enucleated ovum is used to create genetically identical embryos that would be grown in vitro but not be allowed to continue developing to become a human being. The resulting cloned embryos could be used as a source from which to produce stem cells that can then be induced to specialize into the specific type of tissue required by the patient (such as skin for burns victims, brain neuron cells for Parkinson’s disease sufferers, or pancreatic cells for diabetics). The rationale is that because the original nuclear material is derived from a patient’s adult tissue, the risks of rejection of such cells by the immune system are reduced.
6. Gene Therapy
In 1971, Australian Nobel laureate Sir F. MacFarlane Burnet thought that gene therapy (introducing genes into body tissue, usually to treat an inherited genetic disorder) looked more and more like a case of the emperor’s new clothes. Ethical issues aside, he believed that practical considerations forestalled possibilities for any beneficial gene strategy, then or probably ever. Bluntly, he wrote: ‘‘little further advance can be expected from laboratory science in the handling of ‘intrinsic’ types of disability and disease.’’ Joshua Lederberg and Edward Tatum, 1958 Nobel laureates, theorized in the 1960s that genes might be altered or replaced using viral vectors to treat human diseases. Stanfield Rogers, working from the Oak Ridge National Laboratory in 1970, had tried but failed to cure argininemia (a genetic disorder of the urea cycle that causes neurological damage in the form of mental retardation, seizures, and eventually death) in two German girls using Swope papilloma virus. Martin Cline at the University of California in Los Angeles, made the second failed attempt a decade later. He tried to correct the bone marrow cells of two beta-thalassemia patients, one in Israel and the other in Italy. What Cline’s failure revealed, however, was that many researchers who condemned his trial as unethical were by then working toward similar goals and targeting different diseases with various delivery methods. While Burnet’s pessimism finally proved to be wrong, progress in gene therapy was much slower than antibiotic or anticancer chemotherapy developments over the same period of time. While gene therapy had limited success, it nevertheless remained an active area for research, particularly because the Human Genome Project, begun in 1990, had resulted in a ‘‘rough draft’’ of all human genes by 2001, and was completed in 2003. Gene mapping created the means for analyzing the expression patterns of hundreds of genes involved in biological pathways and for identifying single nucleotide polymorphisms (SNPs) that have diagnostic and therapeutic potential for treating specific diseases in individuals. In the future, gene therapies may prove effective at protecting patients from adverse drug reactions or changing the biochemical nature of a person’s disease. They may also target blood vessel formation in order to prevent heart disease or blindness due to macular degeneration or diabetic retinopathy. One of the oldest ideas for use of gene therapy is to produce anticancer vaccines. One method involves inserting a granulocyte-macrophage colony-stimulating factor gene into prostate tumor cells removed in surgery. The cells then are irradiated to prevent any further cancer and injected back into the same patient to initiate an immune response against any remaining metastases. Whether or not such developments become a major treatment modality, no one now believes, as MacFarland Burnet did in 1970, that gene therapy science has reached an end in its potential to advance health.
7. Genetic Engineering
The term ‘‘genetic engineering’’ describes molecular biology techniques that allow geneticists to analyze and manipulate deoxyribonucleic acid (DNA). At the close of the twentieth century, genetic engineering promised to revolutionize many industries, including microbial biotechnology, agriculture, and medicine. It also sparked controversy over potential health and ecological hazards due to the unprecedented ability to bypass traditional biological reproduction.
For centuries, if not millennia, techniques have been employed to alter the genetic characteristics of animals and plants to enhance specifically desired traits. In a great many cases, breeds with which we are most familiar bear little resemblance to the wild varieties from which they are derived. Canine breeds, for instance, have been selectively tailored to changing esthetic tastes over many years, altering their appearance, behavior and temperament. Many of the species used in farming reflect long-term alterations to enhance meat, milk, and fleece yields. Likewise, in the case of agricultural varieties, hybridization and selective breeding have resulted in crops that are adapted to specific production conditions and regional demands. Genetic engineering differs from these traditional methods of plant and animal breeding in some very important respects. First, genes from one organism can be extracted and recombined with those of another (using recombinant DNA, or rDNA, technology) without either organism having to be of the same species. Second, removing the requirement for species reproductive compatibility, new genetic combinations can be produced in a much more highly accelerated way than before. Since the development of the first rDNA organism by Stanley Cohen and Herbert Boyer in 1973, a number of techniques have been found to produce highly novel products derived from transgenic plants and animals.
At the same time, there has been an ongoing and ferocious political debate over the environmental and health risks to humans of genetically altered species. The rise of genetic engineering may be characterized by developments during the last three decades of the twentieth century.
8. Genetic Screening and Testing
The menu of genetic screening and testing technologies now available in most developed countries increased rapidly in the closing years of the twentieth century. These technologies emerged within the context of rapidly changing social and legal contexts with regard to the medicalization of pregnancy and birth and the legalization of abortion. The earliest genetic screening tests detected inborn errors of metabolism and sex-linked disorders. Technological innovations in genomic mapping and DNA sequencing, together with an explosion in research on the genetic basis of disease which culminated in the Human Genome Project (HGP), led to a range of genetic screening and testing for diseases traditionally recognized as genetic in origin and for susceptibility to more common diseases such as certain types of familial cancer, cardiac conditions, and neurological disorders among others. Tests were also useful for forensic, or nonmedical, purposes. Genetic screening techniques are now available in conjunction with in vitro fertilization and other types of reproductive technologies, allowing the screening of fertilized embryos for certain genetic mutations before selection for implantation. At present selection is purely on disease grounds and selection for other traits (e.g., for eye or hair color, intelligence, height) cannot yet be done, though there are concerns for eugenics and ‘‘designer babies.’’ Screening is available for an increasing number of metabolic diseases through tandem mass spectrometry, which uses less blood per test, allows testing for many conditions simultaneously, and has a very low false-positive rate as compared to conventional Guthrie testing. Finally, genetic technologies are being used in the judicial domain for determination of paternity, often associated with child support claims, and for forensic purposes in cases where DNA material is available for testing.
9. Plant Breeding: Genetic Methods
The cultivation of plants is the world’s oldest biotechnology. We have continually tried to produce improved varieties while increasing yield, features to aid cultivation and harvesting, disease, and pest resistance, or crop qualities such as longer postharvest storage life and improved taste or nutritional value. Early changes resulted from random crosspollination, rudimentary grafting, or spontaneous genetic change. For centuries, man kept the seed from the plants with improved characteristics to plant the following season’s crop. The pioneering work of Gregor Mendel and his development of the basic laws of heredity showed for other first time that some of the processes of heredity could be altered by experimental means. The genetic analysis of bacterial (prokaryote) genes and techniques for analysis of the higher (eukaryotic) organisms such as plants developed in parallel streams, but the rediscovery of Mendel’s work in 1900 fueled a burst of activity on understanding the role of genes in inheritance. The knowledge that genes are linked along the chromosome thereby allowed mapping of genes (transduction analysis, conjugation analysis, and transformation analysis). The power of genetics to produce a desirable plant was established, and it was appreciated that controlled breeding (test crosses and back crosses) and careful analysis of the progeny could distinguish traits that were dominant or recessive, and establish pure breeding lines. Traditional horticultural techniques of artificial self-pollination and cross-pollination were also used to produce hybrids. In the 1930s the Russian Nikolai Vavilov recognized the value of genetic diversity in domesticated crop plants and their wild relatives to crop improvement, and collected seeds from the wild to study total genetic diversity and use these in breeding programs. The impact of scientific crop breeding was established by the ‘‘Green revolution’’ of the 1960s, when new wheat varieties with higher yields were developed by careful crop breeding. ‘‘Mutation breeding’’— inducing mutations by exposing seeds to x-rays or chemicals such as sodium azide, accelerated after World War II. It was also discovered that plant cells and tissues grown in tissue culture would mutate rapidly. In the 1970s, haploid breeding, which involves producing plants from two identical sets of chromosomes, was extensively used to create new cultivars. In the twenty-first century, haploid breeding could speed up plant breeding by shortening the breeding cycle.
10. Tissue Culturing
The technique of tissue or cell culture, which relates to the growth of tissue or cells within a laboratory setting, underlies a phenomenal proportion of biomedical research. Though it has roots in the late nineteenth century, when numerous scientists tried to grow samples in alien environments, cell culture is credited as truly beginning with the first concrete evidence of successful growth in vitro, demonstrated by Johns Hopkins University embryologist Ross Harrison in 1907. Harrison took sections of spinal cord from a frog embryo, placed them on a glass cover slip and bathed the tissue in a nutrient media. The results of the experiment were startling—for the first time scientists visualized actual nerve growth as it would happen in a living organism—and many other scientists across the U.S. and Europe took up culture techniques. Rather unwittingly, for he was merely trying to settle a professional dispute regarding the origin of nerve fibers, Harrison fashioned a research tool that has since been designated by many as the greatest advance in medical science since the invention of the microscope.
From the 1980s, cell culture has once again been brought to the forefront of cancer research in the isolation and identification of numerous cancer causing oncogenes. In addition, cell culturing continues to play a crucial role in fields such as cytology, embryology, radiology, and molecular genetics. In the future, its relevance to direct clinical treatment might be further increased by the growth in culture of stem cells and tissue replacement therapies that can be tailored for a particular individual. Indeed, as cell culture approaches its centenary, it appears that its importance to scientific, medical, and commercial research the world over will only increase in the twenty-first century.
History of Biotechnology
Biotechnology grew out of the technology of fermentation, which was called zymotechnology. This was different from the ancient craft of brewing because of its thought-out relationships to science. These were most famously conceptualized by the Prussian chemist Georg Ernst Stahl (1659–1734) in his 1697 treatise Zymotechnia Fundamentalis, in which he introduced the term zymotechnology. Carl Balling, long-serving professor in Prague, the world center of brewing, drew on the work of Stahl when he published his Bericht uber die Fortschritte der zymotechnische Wissenschaften und Gewerbe (Account of the Progress of the Zymotechnic Sciences and Arts) in the mid-nineteenth century. He used the idea of zymotechnics to compete with his German contemporary Justus Liebig for whom chemistry was the underpinning of all processes.
By the end of the nineteenth century, there were attempts to develop a new scientific study of fermentation. It was an aspect of the ‘‘second’’ Industrial Revolution during the period from 1870 to 1914. The emergence of the chemical industry is widely taken as emblematic of the formal research and development taking place at the time. The development of microbiological industries is another example. For the first time, Louis Pasteur’s germ theory made it possible to provide convincing explanations of brewing and other fermentation processes.
Pasteur had published on brewing in the wake of France’s humiliation in the Franco–Prussian war (1870–1871) to assert his country’s superiority in an industry traditionally associated with Germany. Yet the science and technology of fermentation had a wide range of applications including the manufacture of foods (cheese, yogurt, wine, vinegar, and tea), of commodities (tobacco and leather), and of chemicals (lactic acid, citric acid, and the enzyme takaminase). The concept of zymotechnology associated principally with the brewing of beer began to appear too limited to its principal exponents. At the time, Denmark was the world leader in creating high-value agricultural produce. Cooperative farms pioneered intensive pig fattening as well as the mass production of bacon, butter, and beer. It was here that the systems of science and technology were integrated and reintegrated, conceptualized and reconceptualized.
The Dane Emil Christian Hansen discovered that infection from wild yeasts was responsible for numerous failed brews. His contemporary Alfred Jørgensen, a Copenhagen consultant closely associated with the Tuborg brewery, published a widely used textbook on zymotechnology. Microorganisms and Fermentation first appeared in Danish 1889 and would be translated, reedited, and reissued for the next 60 years.
The scarcity of resources on both sides during World War I brought together science and technology, further development of zymotechnology, and formulation of the concept of biotechnology. Impending and then actual war accelerated the use of fermentation technologies to make strategic materials. In Britain a variant of a process to ferment starch to make butadiene for synthetic rubber production was adapted to make acetone needed in the manufacture of explosives. The process was technically important as the first industrial sterile fermentation and was strategically important for munitions supplies. The developer, chemist Chaim Weizmann, later became well known as the first president of Israel in 1949.
In Germany scarce oil-based lubricants were replaced by glycerol made by fermentation. Animal feed was derived from yeast grown with the aid of the new synthetic ammonia in another wartime development that inspired the coining of the word biotechnology. Hungary was the agricultural base of the Austro–Hungarian empire and aspired to Danish levels of efficiency. The economist Karl Ereky (1878–1952) planned to go further and build the largest industrial pig-processing factory. He envisioned a site that would fatten 50,000 swine at a time while railroad cars of sugar beet arrived and fat, hides, and meat departed. In this forerunner of the Soviet collective farm, peasants (in any case now falling prey to the temptations of urban society) would be completely superseded by the industrialization of the biological process in large factory-like animal processing units. Ereky went further in his ruminations over the meaning of his innovation. He suggested that it presaged an industrial revolution that would follow the transformation of chemical technology. In his book entitled Biotechnologie, he linked specific technical injunctions to wide-ranging philosophy. Ereky was neither isolated nor obscure. He had been trained in the mainstream of reflection on the meaning of the applied sciences in Hungary, which would be remarkably productive across the sciences. After World War I, Ereky served as Hungary’s minister of food in the short-lived right wing regime that succeeded the fall of the communist government of Bela Kun.
Nonetheless it was not through Ereky’s direct action that his ideas seem to have spread. Rather, his book was reviewed by the influential Paul Lindner, head of botany at the Institut fu¨ r Ga¨ rungsgewerbe in Berlin, who suggested that microorganisms could also be seen as biotechnological machines. This concept was already found in the production of yeast and in Weizmann’s work with strategic materials, which was widely publicized at that very time. It was with this meaning that the word ‘‘Biotechnologie’’ entered German dictionaries in the 1920s.
Biotechnology represented more than the manipulation of existing organisms. From the beginning it was concerned with their improvement as well, and this meant the enhancement of all living creatures. Most dramatically this would include humanity itself; more mundanely it would include plants and animals of agricultural importance. The enhancement of people was called eugenics by the Victorian polymath and cousin of Charles Darwin, Francis Galton. Two strains of eugenics emerged: negative eugenics associated with weeding out the weak and positive eugenics associated with enhancing strength. In the early twentieth century, many eugenics proponents believed that the weak could be made strong. People had after all progressed beyond their biological limits by means of technology.
Jean-Jacques Virey, a follower of the French naturalist Jean-Baptiste de Monet de Lamarck, had coined the term ‘‘biotechnie’’ in 1828 to describe man’s ability to make technology do the work of biology, but it was not till a century later that the term entered widespread use. The Scottish biologist and town planner Patrick Geddes made biotechnics popular in the English-speaking world. Geddes, too, sought to link life and technology. Before World War I he had characterized the technological evolution of mankind as a move from the paleotechnic era of coal and iron to the neotechnic era of chemicals, electricity, and steel. After the war, he detected a new era based on biology—the biotechnic era. Through his friend, writer Lewis Mumford, Geddes would have great influence. Mumford’s book Technics and Civilization, itself a founding volume of the modern historiography of technology, promoted his vision of the Geddesian evolution.
A younger generation of English experimental biologists with a special interest in genetics, including J. B. S. Haldane, Julian Huxley, and Lancelot Hogben, also promoted a concept of biotechnology in the period between the world wars. Because they wrote popular works, they were among Britain’s best-known scientists. Haldane wrote about biological invention in his far-seeing work Daedalus. Huxley looked forward to a blend of social and eugenics-based biological engineering. Hogben, following Geddes, was more interested in engineering plants through breeding. He tied the progressivism of biology to the advance of socialism.
The improvement of the human race, genetic manipulation of bacteria, and the development of fermentation technology were brought together by the development of penicillin during World War II. This drug was successfully extracted from the juice exuded by a strain of the Penicillium fungus. Although discovered by accident and then developed further for purely scientific reasons, the scarce and unstable ‘‘antibiotic’’ called penicillin was transformed during World War II into a powerful and widely used drug. Large networks of academic and government laboratories and pharmaceutical manufacturers in Britain and the U.S. were coordinated by agencies of the two governments. An unanticipated combination of genetics, biochemistry, chemistry, and chemical engineering skills had been required. When the natural mold was bombarded with high-frequency radiation, far more productive mutants were produced, and subsequently all the medicine was made using the product of these man-made cells. By the 1950s penicillin was cheap to produce and globally available.
The new technology of cultivating and processing large quantities of microorganisms led to calls for a new scientific discipline. Biochemical engineering was one term, and applied microbiology another. The Swedish biologist, Carl-Goran Heden, possibly influenced by German precedents, favored the term ‘‘Biotechnologi’’ and persuaded his friend Elmer Gaden to relabel his new journal Biotechnology and Biochemical Engineering. From 1962 major international conferences were held under the banner of the Global Impact of Applied Microbiology. During the 1960s food based on single-cell protein grown in fermenters on oil or glucose seemed, to visionary engineers and microbiologists and to major companies, to offer an immediate solution to world hunger. Tropical countries rich in biomass that could be used as raw material for fermentation were also the world’s poorest. Alcohol could be manufactured by fermenting such starch or sugar rich crops as sugar cane and corn. Brazil introduced a national program of replacing oil-based petrol with alcohol in the 1970s.
It was not, however, just the developing countries that hoped to benefit. The Soviet Union developed fermentation-based protein as a major source of animal feed through the 1980s. In the U.S. it seemed that oil from surplus corn would solve the problem of low farm prices aggravated by the country’s boycott of the USSR in1979, and the term ‘‘gasohol‘‘ came into currency. Above all, the decline of established industries made the discovery of a new wealth maker an urgent priority for Western governments. Policy makers in both Germany and Japan during the 1970s were driven by a sense of the inadequacy of the last generation of technologies. These were apparently maturing, and the succession was far from clear. Even if electronics or space travel offered routes to the bright industrial future, these fields seemed to be dominated by the U.S. Seeing incipient crisis, the Green, or environmental, movement promoted a technology that would depend on renewable resources and on low-energy processes that would produce biodegradable products, recycle waste, and address problems of the health and nutrition of the world.
In 1973 the German government, seeking a new and ‘‘greener’’ industrial policy, commissioned a report entitled Biotechnologie that identified ways in which biological processing was key to modern developments in technology. Even though the report was published at the time that recombinant DNA (deoxyribonucleic acid) was becoming possible, it did not refer to this new technique and instead focused on the use and combination of existing technologies to make novel products.
Nonetheless the hitherto esoteric science of molecular biology was making considerable progress, although its practice in the early 1970s was rather distant from the world of industrial production. The phrase ‘‘genetic engineering’’ entered common parlance in the 1960s to describe human genetic modification. Medicine, however, put a premium on the use of proteins that were difficult to extract from people: insulin for diabetics and interferon for cancer sufferers. During the early 1970s what had been science fiction became fact as the use of DNA synthesis, restriction enzymes, and plasmids were integrated. In 1973 Stanley Cohen and Herbert Boyer successfully transferred a section of DNA from one E. coli bacterium to another. A few prophets such as Joshua Lederberg and Walter Gilbert argued that the new biological techniques of recombinant DNA might be ideal for making synthetic versions of expensive proteins such as insulin and interferon through their expression in bacterial cells. Small companies, such as Cetus and Genentech in California and Biogen in Cambridge, Massachusetts, were established to develop the techniques. In many cases discoveries made by small ‘‘boutique’’ companies were developed for the market by large, more established, pharmaceutical organizations.
Many governments were impressed by these advances in molecular genetics, which seemed to make biotechnology a potential counterpart to information technology in a third industrial revolution. These inspired hopes of industrial production of proteins identical to those produced in the human body that could be used to treat genetic diseases. There was also hope that industrially useful materials such as alcohol, plastics (biopolymers), or ready-colored fibers might be made in plants, and thus the attractions of a potentially new agricultural era might be as great as the implications for medicine. At a time of concern over low agricultural prices, such hopes were doubly welcome. Indeed, the agricultural benefits sometimes overshadowed the medical implications.
The mechanism for the transfer of enthusiasm from engineering fermenters to engineering genes was the New York Stock Exchange. At the end of the 1970s, new tax laws encouraged already adventurous U.S. investors to put money into small companies whose stock value might grow faster than their profits. The brokerage firm E. F. Hutton saw the potential for the new molecular biology companies such as Biogen and Cetus. Stock market interest in companies promising to make new biological entities was spurred by the 1980 decision of the U.S. Supreme Court to permit the patenting of a new organism. The patent was awarded to the Exxon researcher Ananda Chakrabarty for an organism that metabolized hydrocarbon waste. This event signaled the commercial potential of biotechnology to business and governments around the world. By the early 1980s there were widespread hopes that the protein interferon, made with some novel organism, would provide a cure for cancer. The development of monoclonal antibody technology that grew out of the work of Georges J. F. Kohler and Cesar Milstein in Cambridge (co-recipients with Niels K. Jerne of the Nobel Prize in medicine in 1986) seemed to offer new prospects for precise attacks on particular cells.
The fear of excessive regulatory controls encouraged business and scientific leaders to express optimistic projections about the potential of biotechnology. The early days of biotechnology were fired by hopes of medical products and high-value pharmaceuticals. Human insulin and interferon were early products, and a second generation included the anti-blood clotting agent tPA and the antianemia drug erythropoietin. Biotechnology was also used to help identify potential new drugs that might be made chemically, or synthetically.
At the same time agricultural products were also being developed. Three early products that each raised substantial problems were bacteria which inhibited the formation of frost on the leaves of strawberry plants (ice-minus bacteria), genetically modified plants including tomatoes and rapeseed, and the hormone bovine somatrotropin (BST) produced in genetically modified bacteria and administered to cattle in the U.S. to increase milk yields. By 1999 half the soy beans and one third of the corn grown in the U.S. were modified. Although the global spread of such products would arouse the best known concern at the end of the century, the use of the ice-minus bacteria— the first authorized release of a genetically engineered organism into the environment—had previously raised anxiety in the U.S. in the 1980s.
In 1997 Dolly the sheep was cloned from an adult mother in the Roslin agricultural research institute outside Edinburgh, Scotland. This work was inspired by the need to find a way of reproducing sheep engineered to express human proteins in their milk. However, the public interest was not so much in the cloning of sheep that had just been achieved as in the cloning of people, which had not. As in the Middle Ages when deformed creatures had been seen as monsters and portents of natural disasters, Dolly was similarly seen as monster and as a portent of human cloning.
The name Frankenstein, recalled from the story written by Mary Shelley at the beginning of the nineteenth century and from the movies of the 1930s, was once again familiar at the end of the twentieth century. Shelley had written in the shadow of Stahl’s theories. The continued appeal of this book embodies the continuity of the fears of artificial life and the anxiety over hubris. To this has been linked a more mundane suspicion of the blending of commerce and the exploitation of life. Discussion of biotechnology at the end of the twentieth century was therefore colored by questions of whose assurances of good intent and reassurance of safety could be trusted.
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Biotechnology
The biotech research technique is changing.
All of the ongoing research keeps the potential to bring changes in the quality of life of millions of people, prohibit and do treatment of illnesses that at present have a very high rate of mortality and change healthcare across the world.
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