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Recombinant DNA

About: Recombinant DNA is a(n) research topic. Over the lifetime, 33156 publication(s) have been published within this topic receiving 843935 citation(s). The topic is also known as: rDNA & chimeric DNA. more


Open accessJournal ArticleDOI: 10.1093/NAR/7.6.1513
Abstract: A procedure for extracting plasmid DNA from bacterial cells is described. The method is simple enough to permit the analysis by gel electrophoresis of 100 or more clones per day yet yields plasmid DNA which is pure enough to be digestible by restriction enzymes. The principle of the method is selective alkaline denaturation of high molecular weight chromosomal DNA while covalently closed circular DNA remains double-stranded. Adequate pH control is accomplished without using a pH meter. Upon neutralization, chromosomal DNA renatures to form an insoluble clot, leaving plasmid DNA in the supernatant. Large and small plasmid DNAs have been extracted by this method. more

Topics: Plasmid preparation (74%), Hybrid plasmid (70%), Alkaline lysis (68%) more

13,689 Citations

Open accessJournal ArticleDOI: 10.1128/MCB.2.9.1044
Abstract: We constructed a series of recombinant genomes which directed expression of the enzyme chloramphenicol acetyltransferase (CAT) in mammalian cells. The prototype recombinant in this series, pSV2-cat, consisted of the beta-lactamase gene and origin of replication from pBR322 coupled to a simian virus 40 (SV40) early transcription region into which CAT coding sequences were inserted. Readily measured levels of CAT accumulated within 48 h after the introduction of pSV2-cat DNA into African green monkey kidney CV-1 cells. Because endogenous CAT activity is not present in CV-1 or other mammalian cells, and because rapid, sensitive assays for CAT activity are available, these recombinants provided a uniquely convenient system for monitoring the expression of foreign DNAs in tissue culture cells. To demonstrate the usefulness of this system, we constructed derivatives of pSV2-cat from which part or all of the SV40 promoter region was removed. Deletion of one copy of the 72-base-pair repeat sequence in the SV40 promoter caused no significant decrease in CAT synthesis in monkey kidney CV-1 cells; however, an additional deletion of 50 base pairs from the second copy of the repeats reduced CAT synthesis to 11% of its level in the wild type. We also constructed a recombinant, pSV0-cat, in which the entire SV40 promoter region was removed and a unique HindIII site was substituted for the insertion of other promoter sequences. more

Topics: Chloramphenicol acetyltransferase (59%), Molecular cloning (54%), Recombinant DNA (53%) more

7,430 Citations

08 Apr 1983-
Abstract: Altered and native immunoglobulins, including constant-variable region chimeras, are prepared in recombinant cell culture. The immunoglobulins contain variable regions which are immunologically capable of binding predetermined antigens. Methods are provided for refolding directly expressed immunoglobulins into immunologically active form. more

Topics: Recombinant DNA (51%)

5,955 Citations

Journal ArticleDOI: 10.1126/SCIENCE.322279
WD Benton1, Ronald W. Davis1Institutions (1)
08 Apr 1977-Science
Abstract: A rapid, direct method for screening single plaques of Agt recombinant phage is described. The method allows at least 10(6) clones to be screened per day and simplifies physical containment of recombinants. more

Topics: Recombinant DNA (52%)

3,920 Citations

PatentDOI: 10.1073/PNAS.95.5.2509
Abstract: Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. This invention describes a strategy which simplifies the generation and production of such viruses. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, employing homologous recombination in bacteria rather than in eucaryotic cells. Following transfections of such plasmids into a mammalian packaging cell line, viral production can be conveniently followed with the aid of green fluorescent protein, encoded by a gene incorporated into the viral backbone. Homogeneous viruses can be obtained from this procedure without plaque purification. This system expedites the process of generating and testing recombinant adenoviruses. more

Topics: Recombinant DNA (51%), Plasmid (51%)

3,509 Citations

No. of papers in the topic in previous years

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Topic's top 5 most impactful authors

Florian M. Wurm

36 papers, 1.8K citations

Mathias Uhlén

30 papers, 843 citations

Bernard Moss

24 papers, 1.9K citations

Pierre Tiollais

19 papers, 2.1K citations

Ira Pastan

16 papers, 997 citations

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