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PubMed1983

Human insulin from recombinant DNA technology.

Human insulin produced by recombinant DNA technology is the first commercial health care product derived from this technology. Work on this product was initiated before there were federal guidelines for large-scale recombinant DNA work or commercial development of recombinant DNA products. The steps taken to facilitate acceptance of large-scale work and proof of the identity and safety of such a product are described. While basic studies in recombinant DNA technology will continue to have a profound impact on research in the life sciences, commercial applications may well be controlled by economic conditions and the availability of investment capital.

Cloning, Molecular↗
PubMed1985

From DNA to NDA--the impact of recombinant DNA technology on new drug development.

Man's long-standing efforts to alter living things through genetic manipulation have become reality. Recent advances in recombinant DNA technology have the potential to alter the drug-development process profoundly. The pharmaceutical industry has had to adjust its research efforts and develop new state-of-the-art laboratories. In addition to the standard biological and in vivo assays, many new tests are required, e.g., amino acid sequencing, high-pressure liquid chromatography, and radioimmunoassays. Academic researchers have played a vital role in developing the new biotechnology, supplying most of the basic scientific knowledge and the initial supply of the scientific work force. The recent shifting of support for scientific training from the government to the pharmaceutical industry has resulted in unprecedented academe-industry relationships. Universities now stand to profit significantly from patent rights resulting from biotechnology research efforts. While the advances in biotechnology have had considerable impact on the pharmaceutical industry and academia, they have thus far had only a minor impact on the regulatory process. To date, the preferred regulatory path appears to be modification of existing procedures through the issuance of guidelines, which can be updated as knowledge increases.

Academies and Institutes↗
PubMed2003

The role of DNA recombination in herpes simplex virus DNA replication.

In many organisms the processes of DNA replication and recombination are closely linked. For instance, in bacterial and eukaryotic systems, replication forks can become stalled or damaged, in many cases leading to the formation of double stranded breaks. Replication restart is an essential mechanism in which the recombination and repair machinery can be used to continue replication after such a catastrophic event. DNA viruses of bacteria such as lambda and T4 also rely heavily on DNA recombination to replicate their genomes and both viruses encode specialized gene products which are required for recombination-dependent replication. In this review, we examine the linkage between replication and recombination in the eukaryotic pathogen, Herpes Simplex Virus Type 1 (HSV-1). The evidence that recombination plays an intrinsic role in HSV-1 DNA replication and the infection process will be reviewed. We have recently demonstrated that HSV-1 encodes two proteins which may be analogous to the lambda phage recombination system, Red(alpha) and beta. The HSV-1 alkaline nuclease, a 5' to 3' exonuclease, and ICP8, a single stranded DNA binding protein, can carry out strand annealing reactions similar to those carried out by the lambda Red system. In addition, evidence suggesting that host recombination proteins may also be important for HSV-1 replication will be reviewed. In summary, it is likely that HSV-1 infection will require both viral and cellular proteins which participate in various pathways of recombination and that recombination-dependent replication is essential for the efficient replication of viral genomes.

Bacteria↗
PubMed1990

DNA based diagnostic tests: recombinant DNA and cardiovascular disease risk factors.

Advances in molecular biology and medical biotechnology are continuously creating exciting possibilities for DNA based diagnostics. It is now possible by simple procedures to detect polymorphic DNA markers, structural variants and regulatory mutants of human genes, allowing detailed genotyping of patients. The innovative combination of immunoenzymatic techniques, monoclonal antibodies and recombinant tracer proteins, results in new DNA based tests for the determination of important biochemical parameters, in order to define more precisely the phenotype and hence assess the individual risk. The application of these technologies to the analysis of dyslipidemias, atherosclerosis and cardiovascular diseases may not only lead to a better understanding of the molecular and genetic basis of these pathologies, but also to their early recognition and better management.

Base Sequence↗
PubMed1985

Applications of recombinant DNA to pathologic diagnosis.

Recombinant DNA techniques are contributing to the understanding of the pathogeneses of genetic, neoplastic, and viral diseases, and are used in the diagnosis of certain genetic and viral diseases. Such techniques will have wider application in the future and will play an increasing role in the clinical laboratory. The technology of this field rests upon the cleavage of DNA by certain enzymes, restriction endonucleases, and upon the ability to locate specific sequences of nucleotides in a cleaved DNA sample by using known fragments of DNA ("probes") labeled with radioisotopes or biotin. To produce useful probes, one "clones" multiple copies of the same DNA fragment in bacteria. The use of DNA probes in the clinical laboratory is valuable in antenatal diagnosis, genetic counseling, and post-natal diagnosis of genetic diseases, especially hematologic diseases and inborn errors of metabolism. DNA probes can also be used to detect viral genetic material in clinical specimens.

Adult↗
PubMedDate not supplied

Less pronounced changes in serum potassium and epinephrine during hypoglycemia induced by human insulin (recombinant DNA).

Human insulin (recombinant DNA) and purified porcine insulin were compared in healthy men. Intravenous insulin tolerance tests showed identical effects on plasma glucose when a bolus of 0.1 U/kg was injected. Following human insulin, hypokalemia and epinephrine secretion were significantly less pronounced. The differences in serum potassium concentrations are caused by a lower epinephrine response to hypoglycemia induced by human insulin in comparison to purified porcine insulin.

Adult↗
PubMed1986

Environmental implications of recombinant DNA technology.

Applications of recombinant DNA technology are discussed as a backdrop for evaluation of the environmental impacts of this technology. Some of applications include using traditional biological techniques for specific purposes, including nitrogen fixation, microbial pesticides, and waste treatment. In these instances the final product lies along a continuum, beginning with an organism marginally performing its function, and ending with one that is highly specialised and very efficient in what it does. One may move along this continuum toward the 'perfect' microorganism by using traditional methodologies of mutagenesis and selection, recombinant DNA technology, or a combination of the two.

Journal Article↗
PubMed1990

Bcr/abl recombinant DNA analysis versus karyotype in the diagnosis and therapeutic monitoring of chronic myeloid leukemia.

Karyotype and bcr/abl recombinant DNA analyses are two means of detecting the chromosomal aberration in chronic myeloid leukemia. The authors compared these two methods in a retrospective study of 36 patients with CML in which they found the bcr/abl DNA recombinant event in 100% (29 of 29) of those patients who had the Philadelphia chromosome. To achieve this sensitivity, a battery of two bcr probes and three restriction enzymes is necessary. The authors propose a sequential algorithm for efficient use of these probes and enzymes. In 76% of the patients, bcr/abl rearrangement can be detected with a Bgl II digest and a 3' commercial probe. An additional 21% of patients can be detected by a second assay in which the same membrane is rehybridized to a 3' and 5' combination bcr probe. One patient (3%) required an additional restriction enzyme digest with BamH I to detect the recombinant event by the same 3' probe. Karyotype analysis is used to determine cytogenetic remission in patients with CML under therapy. The authors studied the use of DNA analysis by the Southern blot technique to detect a decrease in the relative number of leukemic cells. By dilution studies and densitometric scanning of autoradiographs, the authors were able to detect a 15% decrease in the relative number of cells having the bcr/abl recombinant event. The authors report the preliminary results of three patients in whom they compared the karyotype and recombinant DNA analysis at multiple time points in their clinical course. In conclusion, the bcr/abl recombinant DNA analysis is superior to karyotype for the diagnosis of CML and can be used for monitoring treated patients.

Adolescent↗
PubMed1995

The production of K88 antigen by Escherichia coli and Salmonella typhimurium strains with recombinant DNA.

Expression of K88ab antigen by strains with recombinant DNA, differing in molecular weight or promoter, was subjected to investigation. Strains with recombinant DNA produced greater amounts of antigen than did field isolates. Maximal production was recorded for the E. coli C600 strain with 10.85 kb recombinant DNA with promoter P1 of the pBR322 vector. The substitution of promoter Ptac for promoter P1 did not result in an increased expression of K88ab antigen. The production of K88ab antigen by Salmonella typhimurium TM333 with recombinant DNA was on a level comparable to that of E. coli C600 or E. coli HB101 strains with the same recombinant DNA.

Animals↗
PubMed1986

A cell hybrid and recombinant DNA library that facilitate identification of polymorphic loci in the vicinity of the Huntington disease gene.

Somatic cell hybrids were selected that retain a derivative chromosome 5 from an individual in which the p15.1-pter segment of chromosome 5 is replaced with the p15.1-pter segment of chromosome 4. Hybrids that retain this derivative chromosome exclusively were found to be positive for G8, a DNA marker closely linked to the Huntington disease gene on chromosome 4p. From one such hybrid, a segregant was isolated that had deleted the entire q arm of the derivative chromosome but retained the p arm intact as its only detectable human DNA. A complete recombinant DNA library was prepared from this cell line, and the inserts in approximately 1/3 of the recombinant phage with human DNA were shown to be derived from 4pter-4p15.1, which represents only approximately 1% of the total human genome. The cell hybrid and DNA library represent a rapid and efficient means to identify and isolate many polymorphic DNA markers close to and flanking the Huntington disease gene.

Animals↗
PubMed1987

Recombinant DNA in filamentous fungi: progress and prospects.

Recombinant DNA technology enables the creation of well-defined alterations in the genetic material of an organism. Methods to manipulate recombinant DNA in the filamentous fungi (a group of microorganisms that includes species of academic as well as commercial interest) have recently been developed. This has been the result of adaptation of procedures successfully employed in the manipulation of other microorganisms. There are a number of similarities in the behavior of recombinant DNA in different fungi, but a number of differences have also been observed between the filamentous and the nonfilamentous fungi. Such differences include the ability to identify DNA replication origins and the host range of expression of fungal genes.

Cloning, Molecular↗