PCR amplification of large genomic fragments from human and simian immunodeficiency virus infected cell lines.
Polymerase chain reaction (PCR) has been used to amplify the large fragments from viral genomic DNA of SIV from wild caught, asymptomatic 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.
Saksena
N K
NK
SUNY, HSC. Department of Medicine, Syracuse 13203.
Dwyer
D E
DE
Müller
M
M
Durand
J P
JP
Hervé
V
V
Lang
M C
MC
Barré-Sinoussi
F
F
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D016384:Consensus Sequence; D016669:Erythrocebus; D015752:Genes, env; D015750:Genes, gag; D015751:Genes, pol; D016679:Genome, Viral; D015498:HIV-2 / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D015302:Simian Immunodeficiency Virus / Q000235:genetics* / Q000254:growth & development / Q000648:ultrastructure
D009838:Oligodeoxyribonucleotides; D012194:RNA-Directed DNA Polymerase