PCR amplification of large genomic fragments from human and simian immunodeficiency virus infected cell lines
| Autor: | Nitin K. Saksena, Vincent Herve, Michaela C. Muller, Marie-Claude Lang, Jean Paul Durand, Dominic E. Dwyer, Françoise Barré-Sinoussi |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
viruses
Molecular Sequence Data Genome Viral Biology medicine.disease_cause Genes env Polymerase Chain Reaction Genome General Biochemistry Genetics and Molecular Biology Cell Line law.invention chemistry.chemical_compound law Consensus Sequence medicine Consensus sequence Animals Humans Erythrocebus General Pharmacology Toxicology and Pharmaceutics Gene Polymerase chain reaction Base Sequence RNA-Directed DNA Polymerase General Medicine Simian immunodeficiency virus Genes gag Genes pol Virology Molecular biology Long terminal repeat Blotting Southern Microscopy Electron genomic DNA Oligodeoxyribonucleotides chemistry HIV-2 Simian Immunodeficiency Virus DNA |
| Zdroj: | Life Sciences. 50:1973-1984 |
| ISSN: | 0024-3205 |
| Popis: | Polymerase chain reaction (PCR) has been used to amplify the large fragments from viral genomic DNA of SIV from wild caught, asymptotic Erythrocebus monkeys from Western Africa (Senegal) and also from HIV-2 infected cell lines. By using consensus primer sequences from highly conserved stretches of gag, pol and env genes, two halves of the viral genome of HIV-2 and SIV (isolated from west African Erythrocebus monkeys) have amplified by PCR. One half spans 5200 bp from within the U3 region of the 5′ long terminal repeat (LTR) into pol gene and an overlapping fragment spans 3700 bp from the pol gene into U5 region of 3′ LTR. Also fragments ranging from 1–2.3 kb from gag pol and env genes have been successfully amplified. Our data demonstrate that primers used to amplify large segments from viral DNA yield better results if they are derived from a consensus sequence of a highly conserved stretch of the viral genome. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|