Homologous recombination between human immunodeficiency viral DNAs in cultured human cells: analysis of the factors influencing recombination.
Recombination between HIV DNAs was analyzed using DNA transfection in cell cultures and the optimal conditions for efficient recombination were determined. Recombinant plasmid DNA substrates were constructed from HIV proviral DNAs and the success of recombination was measured by the production of viable hybrid virus. The process of recombination between HIV DNAs was shown to be i) dependent on homology between the truncated HIV DNAs and ii) maximum with concentrations of the truncated DNAs 3ug and above. HIV isolates with heterogeneity in their primary sequence, thus offer an ideal system for the analysis of the requirement of homologous recombination. In addition, recombination methodology would be useful for generating hybrid HIVs for the analysis of specific viral gene functions.
Kalyanaraman
S
S
Retrovirus Diseases Branch Centers for Disease Control, Atlanta, Georgia 30333.
Jannoun-Nasr
R
R
York
D
D
Luciw
P A
PA
Robinson
R
R
Srinivasan
A
A
D002130:Calcium Phosphates / Q000378:metabolism; D002452:Cell Count; D002528:Cerebellar Neoplasms; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D005814:Genes, Viral; D014644:Genetic Variation; D006678:HIV / Q000235:genetics* / Q000502:physiology; D006801:Humans; D008527:Medulloblastoma; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase; D011995:Recombination, Genetic; D012208:Rhabdomyosarcoma; D012689:Sequence Homology, Nucleic Acid; D014162:Transfection; D014407:Tumor Cells, Cultured; D014779:Virus Replication
D002130:Calcium Phosphates; D004274:DNA, Recombinant; D004279:DNA, Viral; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; C485838:calcium phosphate, monobasic, anhydrous; C020243:calcium phosphate; D012194:RNA-Directed DNA Polymerase; C485829:calcium phosphate, dibasic, anhydrous