Optimal conditions for synthesizing complementary DNA in the HIV-1 endogenous reverse transcriptase reaction.
Concentration of monovalent and divalent cations, anionic detergent, reducing agent and nucleotides, as well as pH, temperature, and incubation time were optimized for high levels of HIV-1 endogenous reverse transcriptase activity. In addition, mellitin, a peptide substitute for anionic detergent, and oligo(dT)12-18 were found to stimulate nucleic acid synthesis. This HIV-1 endogenous reaction demonstrated RNA- and DNA-dependent DNA polymerase activities. Nucleic acid intermediates and final products included RNA:DNA hybrids as well as single- and double-stranded DNA. The complementary DNA products formed were representative of all regions of the HIV-1 genome.
Cancer Research Institute, University of California School of Medicine, San Francisco 94143-0128.
W H
WH
Yong
S
S
Wyman
J A
JA
Levy
D002412:Cations; D004247:DNA / Q000096:biosynthesis* / Q000235:genetics; D005814:Genes, Viral; D005821:Genetic Techniques; D015497:HIV-1 / Q000235:genetics*; D007202:Indicators and Reagents; D008555:Melitten; D009693:Nucleic Acid Hybridization; D009838:Oligodeoxyribonucleotides; D012194:RNA-Directed DNA Polymerase; D014158:Transcription, Genetic
D002412:Cations; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; C027903:oligo (dT); D008555:Melitten; D004247:DNA; D012194:RNA-Directed DNA Polymerase