Generation of hybrid human immunodeficiency virus by homologous recombination.
Human immunodeficiency virus (HIV) type 1, isolated from diverse sources, exhibits genomic diversity. The mechanisms by which the genomic diversity takes place in individuals exposed to multiple virus isolates is yet to be elucidated. Genetic variation, in general, might result from mutagenic events such as point mutations, rearrangements (insertions and deletions), and recombination. In an attempt to evaluate the process of genetic diversity, we designed experiments to analyze recombination between HIV DNAs by using DNA transfection in cell cultures. Here we report the successful recombination between truncated HIV proviral DNAs with an overlap homology of 53 base pairs that leads to the formation of viable hybrid virus. Recombination was also seen between exogenous DNA introduced into cells and homologous HIV sequences resident in the cells. These results indicate that recombination among various HIV isolates may play a significant role in the generation of genetic diversity of HIV. Further, the method used here enables the construction of hybrid HIV genomes to identify the viral determinants responsible for tropism, replication, and cytopathic effects.
Srinivasan
A
A
Retrovirus Diseases Branch, Centers for Disease Control, Atlanta, GA 30333.
York
D
D
Jannoun-Nasr
R
R
Kalyanaraman
S
S
Swan
D
D
Benson
J
J
Bohan
C
C
Luciw
P A
PA
Schnoll
S
S
Robinson
R A
RA
D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D015488:HIV Antigens / Q000032:analysis; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase / Q000032:analysis; D011995:Recombination, Genetic; D015183:Restriction Mapping; D014162:Transfection
D004279:DNA, Viral; D015488:HIV Antigens; D012194:RNA-Directed DNA Polymerase