PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Genetic Engineering”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 73 records · Page 4Linked to original sources
PubMed1982

Genetically engineered viral vaccines--prospects for the future.

Genetic engineering (recombinant DNA technology)--the revolution in molecular biology--has enabled us to isolate any genes from any source in a pure form, and to move them from one cell to another. It has become possible to program bacterial or yeast cells with foreign genes and force the new host to produce commercially valuable proteins (e.g. hormones, enzymes, diagnostic reagents). It is now also possible to produce viral and bacterial antigens in various types of cells. We hope that this will soon enable us to manufacture vaccines cheaply. The production of a foot-and-mouth-disease virus vaccine--the first promising example of a genetically engineered effective vaccine--has recently been reported. Expression of hepatitis B surface antigen, influenza virus haemagglutinin and polio-virus proteins from the cloned genes have also been reported, and many more viral genes have been cloned although not yet expressed in bacteria. Despite the extremely rapid development, there are a number of problems, both technical and immunological, which have to be extensively studied and eventually solved, before we can hope to obtain effective and safe genetically engineered viral vaccines for clinical use.

Bacteria↗
PubMed2001

Bioluminescence immunoassay for thyroxine employing genetically engineered mutant aequorins containing unique cysteine residues.

Genetically engineered one-to-one conjugates between an analyte and a protein label have been demonstrated to yield assays with better detection limits and performance characteristics than those prepared by conventional chemical conjugation methods. To date, the preparation of these conjugates has been limited to fusion techniques where a peptide analyte is fused in frame to the protein label. To further expand the range of analytes that can be detected by using genetic engineering techniques coupled with bioanalytical methods, we have employed site-directed mutagenesis to prepare one-to-one analyte-label conjugates that include nonpeptidic analytes such as drugs, vitamins, and hormones. Specifically, we have prepared mutants of the photoprotein aequorin containing single cysteine residues suitable for site-specific conjugation. Aequorin is a photoprotein that emits light at 469 nm and has been employed as a highly sensitive bioluminescent label in the development of binding assays for important biomolecules. We have performed polymerase chain reaction-based site-directed mutagenesis on apoaequorin to yield four mutant aequorins containing unique cysteine residues at positions 5, 53, 71, and 84 in the polypeptide chain for the purpose of site-specific conjugation to a model analyte. A maleimide-activated thyroxine was selected as the model analyte and site-specifically conjugated to the mutants through their unique cysteine residues. A heterogeneous assay for thyroxine was then developed by employing the genetically engineered aequorin mutants.

Aequorin↗
PubMedDate not supplied

[Bioethical considerations of genetic engineering].

Recombinant DNA technology, popularly known as genetic engineering, has numerous applications which have both fascinated and alarmed the public. Since the mid-70s, the benefits of this technology, generally speaking, outnumber the risks. These benefits, which take the shape of an increase in knowledge and in the improvement of diagnostic and therapeutic products, have had widespread repercussions, not only in the laboratory but also in our day-to-day lives. Scientific progress and the new scientific directions in Biology make it necessary for scientists to have an advanced philosophical concept of the world so as to be able to properly evaluate the results and perspectives of any biological science, including those of genetic engineering. It is very important for it to have a biosocial approach: there must be an unrelenting battle against pseudoscientific ideas and attempts to use the recent advances in Biology for reactionary, antihuman purposes.

Bioethical Issues↗
PubMed2000

Genetic engineering of single-chain gonadotropins and hormone-receptor fusion proteins.

The gonadotropin hormone family is distinguished by its heterodimeric structure in which the members share a common alpha subunit and a hormone-specific beta subunit. Since assembly of the heterodimer is often the rate-limiting step in production of functional hormone, single-chain hormones have been engineered by genetically linking the two subunits. The single-chain hormone can in turn be fused to its receptor to produce a functional single-chain hormone-receptor complex. These fusion constructs offer a valuable new approach in structure-function studies and in the generation of hormone analogs. In this article we describe the experimental design for the generation of single-chain human chorionic gonadotropin and single-chain hormone-receptor fusion complex and strategies for the expression of these fusion proteins.

Amino Acid Sequence↗
PubMed1993

[New vaccines in the age of genetic engineering].

The era of genetic engineering is merely 20 years of age, yet has already borne completely new perspectives for vaccine development. Important insights are gained by elucidating the genetic information of a disease-causing microorganism and its pathogenic and attenuated variants. Site-directed mutagenesis can then be employed to specifically alter the genetic information in a variety of ways. Upon transfer of the corresponding gene to pro- or eucaryotic cells, large quantities of microbial components can be produced. A new generation of such subunit vaccines is already undergoing clinical testing. Recently, hybrid vaccines have been constructed, which utilize highly successful traditional live vaccines such as polio- and vaccinia-virus or the Tbc-bacterium as carriers for components of other microorganisms. We should bear in mind that the same new technologies can be abused for the construction of potentially dangerous biological weapons. The scientific community bears the responsibility to prevent such abuse and to lobby for an easy access to the new vaccines by the world's poorest inhabitants.

Bacterial Vaccines↗
PubMed1999

Genome system architecture and natural genetic engineering in evolution.

Molecular genetics teaches three lessons relevant to the nature of genetic change during evolution: (1) Genomes are organized as hierarchies of composite systems (multidomain protein-coding sequences; functional loci made up of regulatory, coding, processing, and intervening sequences; and multilocus regulons and replicons) interconnected and organized into specific "system architectures" by repetitive DNA elements. (2) Genetic change often occurs via natural genetic engineering systems (cellular biochemical functions, such as recombination complexes, topoisomerases, and mobile elements, capable of altering DNA sequence information and joining together different genomic components). (3) The activity of natural genetic systems is regulated by cellular control circuits with respect to the timing, activity levels, and specificities of DNA rearrangements (e.g., adaptive mutation, Ty element mobility, and P factor insertions). These three lessons provide plausible molecular explanations for the episodic, multiple, nonrandom DNA rearrangements needed to account for the evolution of novel genomic system architectures and complex multilocus adaptations. This molecular genetic perspective places evolutionary change in the biologically responsive context of cellular biochemistry.

Adaptation, Biological↗
PubMed2006

The experimental study of genetic engineering human neural stem cells mediated by lentivirus to express multigene.

OBJECTIVE: To explore the feasibility to construct genetic engineering human neural stem cells (hNSCs) mediated by lentivirus to express multigene in order to provide a graft source for further studies of spinal cord injury (SCI). METHODS: Human neural stem cells from the brain cortex of human abortus were isolated and cultured, then gene was modified by lentivirus to express both green fluorescence protein (GFP) and rat neurotrophin-3 (NT-3); the transgenic expression was detected by the methods of fluorescence microscope, dorsal root ganglion of fetal rats and slot blot. RESULTS: Genetic engineering hNSCs were successfully constructed. All of the genetic engineering hNSCs which expressed bright green fluorescence were observed under the fluorescence microscope. The conditioned medium of transgenic hNSCs could induce neurite flourishing outgrowth from dorsal root ganglion (DRG). The genetic engineering hNSCs expressed high level NT-3 which could be detected by using slot blot. CONCLUSIONS: Genetic engineering hNSCs mediated by lentivirus can be constructed to express multigene successfully.

Animals↗
PubMed1989

Genetically Engineered Erwinia carotovora in Aquatic Microcosms: Survival and Effects on Functional Groups of Indigenous Bacteria.

The survival of genetically engineered Erwinia carotovora L-864, with a kanamycin resistance gene inserted in its chromosome, was monitored in the water and sediment of aquatic microcosms. The density of genetically engineered and wild-type E. carotovora strains declined at the same rate, falling in 32 days below the level of detection by viable counts. We examined the impact of the addition of genetically engineered and wild-type strains on indigenous bacteria belonging to specific functional groups important in nutrient cycling. For up to 16 days, the densities of total and proteolytic bacteria were significantly higher (P < 0.05) in microcosms inoculated with genetically engineered or wild-type E. carotovora, but by 32 days after inoculation, they had decreased to densities similar to those in control microcosms. Inoculation of genetically engineered or wild-type E. carotovora had no apparent effect on the density of amylolytic and pectolytic bacteria in water and sediment. Genetically engineered and wild-type E. carotovora did not have significantly different effects on the densities of specific functional groups of indigenous bacteria (P > 0.05).

Journal Article↗
PubMed2006

Effect of ambient conditions on simultaneous growth and bioaccumulation of mercuric ion by genetically engineered E. coli JM109.

Genetically engineered E. coli JM109, namely M1, which expressed both Hg(2+) transport system and metallothionein, was tested for its capability of simultaneous growth and bioaccumulation of Hg(2+) under low nutritional circumstances. The influential factors of ambient conditions, e.g. initial concentrations of mercuric ion, ionic strength, the presence of metal chelators and other coexisting metal ions were investigated. Hg(2+) bioaccumulation behavior of M1 proved to be well coupled with its growth. NaCl was essential to the growth of M1. Of all tested NaCl concentrations, 0.04 mol/L was optimal. The presence of 0.1 mol/L CaCl(2) or MgCl(2) could promote the growth of M1 and keep the Hg(2+) removal ratio high, but the growth of M1 was inhibited seriously as the concentration of CaCl(2) or MgCl(2) reached 0.3 mol/L. Chelator EDTA had a significant influence on M1 growth and Hg(2+) bioaccumulation, while the effect of citration was little. The presence of other coexisting metal ions inhibited the growth of M1. The influential order was as follows: Cd(2+)>Zn(2+)> or =Cu(2+)>Pb(2+)>Ni(2+). However, only Cd(2+) and Cu(2+) posed obviously adverse effects on Hg(2+) bioaccumulation during the SG&B process.

Chelating Agents↗
PubMed2004

How against HIV - using immunology or genetic engineering.

How against HIV - using Immunology or Genetic Engineering represents a thinking about the way how to overarch the HIV virus. Namely, by the combination of Immunological and the methods of Genetic Engineering, we could create a new nonreproducible virus which would be a competitor to real HIV for CD4 receptors. Competitors, by their affinity, would be acting in order to engage the CD4 receptors, making them nonaccessible for real HIV. But, at the same time, competitors, by their size, would be just physically covering a small deal of CD4 receptors. That way these free CD4 receptors would also be nonaccessible for real HIV, but accessible for immunomolecules, like, for an example MHC II is. That way the physiological role of CD4 receptors in immunological reactions, would be saved, and that fact would help us in our intention to expel the provirus integrated in its host CD4 lymphocite, by already known immunological mechanisms. Out of the host cell (CD4 Lymphocite) the HIV virus is very responsive for our Immune System and it would be destroyed.

Acquired Immunodeficiency Syndrome↗
PubMedDate not supplied

Site-specific conjugation of a temperature-sensitive polymer to a genetically-engineered protein.

A genetically-engineered mutant of cytochrome b5, incorporating a unique cysteine residue, was conjugated to maleimide-terminated oligo(N-isopropylacrylamide). The conjugation of the protein by reaction of the cysteine residue, precisely positioned by site-directed mutagenesis techniques, with an activated oligomer containing only one reactive end group in the oligomer chain permits the site-specific and stoichiometric conjugation of the oligomer with the protein. The protein-oligomer conjugate was shown to exhibit lower critical solution temperature (LCST) behavior, similar to the free oligomer. Furthermore, the LCST behavior of the protein-oligomer conjugate is reversible and allows selective precipitation of the conjugate above its LCST.

Acrylic Resins↗
PubMed2001

Procedures for microencapsulation of enzymes, cells and genetically engineered microorganisms.

Methods to microencapsulate enzyme, cells, and genetically engineered cells have been described in this article. More specific examples of enzyme encapsulation include the microencapsulation of xanthine oxidase for Lesch-Nyhan disease; phenylalanine ammonia lyase for pheny, ketonuria and microencapsulation of multienzyme systems with cofactor recycling for multistep enzyme conversions. Methods for cell encapsulation include the details for encapsulating hepatocytes for liver failure and for gene therapy. This also includes the details of a novel two-step method for encapsulation of high concentrations of smaller cells. Another new approach is the detailed method of the encapsulation of genetically engineered Escherichia coli DH5 cells for lowering urea, ammonia, and other metabolites in kidney or, liver failure and other diseases.

Animals↗
PubMed2006

Use of a risk communication model to evaluate dietetics professionals' viewpoints on genetically engineered foods and crops.

The complex issues surrounding the application of genetic engineering to food and agriculture have generated a contentious debate among diverse interest groups. One pervasive dimension in the resultant discourse is the varying perceptions of the risks and benefits of genetically engineered foods and crops. In the risk communication model, technical information is evaluated within the context of an individual's values and perceptions. The purpose of this study was to explore how dietetics professionals respond to a complex set of interrelated issues associated with genetically engineered foods and crops and to identify what varying viewpoints may exist. Participants were asked to sort a total of 48 statements distributed across eight issue areas according to level of agreement and disagreement. Using Q methodology, a total of 256 sortings were analyzed using the centroid method and varimax rotation in factor analysis. Three distinct viewpoints emerged: Precautionary (R(2)=43%), Discerning Supporter (R(2)=11%), and Promoting (R(2)=5%). Across all viewpoints, respondents agreed that dietetics professionals should employ critical thinking skills to communicate the social, economic, environmental, ethical, and technical aspects of genetically engineered foods and crops. The findings have implications for how dietetics professionals can foster an open interchange of information among diverse groups.

Attitude of Health Personnel↗
PubMed1993

Improvement of downstream processing of recombinant proteins by means of genetic engineering methods.

The rapid advancement of genetic engineering has allowed to produce an impressive number of proteins on a scale which would not have been achieved by classical biotechnology. At the beginning of this development research was focussed on elucidating the mechanisms of protein overexpression. The appearance of inclusion bodies may illustrate the success. In the meantime, genetic engineering is not only expected to achieve overexpression, but to improve the whole process of protein production. For downstream processing of recombinant proteins, the synthesis of fusion proteins is of primary importance. Fusion with certain proteins or peptides may protect the target protein from proteolytic degradation and may alter its solubility. Intracellular proteins may be translocated by means of fusions with signal peptides. Affinity tags as fusion complements may render protein separation and purification highly selective. These methods as well as similar ones for improving the downstream processing of proteins will be discussed on the basis of recent literature.

Journal Article↗
PubMed1985

Genetically engineered insulin: five years of experience.

Genetically engineered insulin is the first application of recombinant DNA technology which has gone into industrial production and wide clinical use. Four years after the first clinical trials, it appears that there are only minor pharmacokinetic differences from purified pork insulin; in particular, a faster subcutaneous absorption for both regular and NPH forms. The hypoglycaemic potency of genetically engineered insulin is identical to that of purified pork insulin but a weaker effect on counterregulatory hormones has been reported. However, the main advantage of biosynthetic human insulin is its species specificity, which reduces its immunogenicity. Convincing results were obtained in patients suffering from insulin-produced adverse reactions such as insulin resistance or allergy, although biosynthetic human insulin does have some immunological properties and crossreacts with beef or pork insulin antibodies.

Animals↗
PubMed1994

Survival and impact of genetically engineered Pseudomonas putida harboring mercury resistance gene in soil microcosms.

The survival of genetically engineered and wild-type Pseudomonas putida PpY101, that contained a recombinant plasmid pSR134 conferring mercury resistance, were monitored in andosol and sand microcosms. The survival of genetically engineered and wild-type P. putida was not significantly different in andosol. The population change of the two strains was dissimilar in andosol and sand. The survival of genetically engineered and wild-type P. putida strains was affected by the water content of andosol, and increased with the increment of the water content. The impact of the addition of genetically engineered and wild-type P. putida strains on indigenous bacteria and fungi was examined. Inoculation of both strains had no apparent effect on the density of indigenous microorganisms.

Drug Resistance, Microbial↗