关键词: TRIM5α, HIV-1, 病毒

Abstract: Objective To study the function of anti-HIV activity of TRIM5α.
Methods The full-length cDNA encoding TRIM5α in American green monkey(AGM) was obtained using the reverse transcription polymerase chain reaction(RT-PCR) method after extracting the total RNA from vero cells. The cDNA fragments corresponding to the B30.2 and coiled coil region of TRIM5α from human, rhesus monkey, and AGM were amplified with routine PCR method and ligated into the expression vector pEGFP-C3. After fusion expression with green fluorescence, the cellular location of the protein was observed under fluorescent microscope. Pseudotyped viruses were generated by co-transfection of HIV Δenv-expressing plasmid and VSV-G-expressing plasmid. The anti-HIV activity of TRIM5α and its fragments were quantitatively tested by the inhibition of pseudotyped virus infection.
Results The plasmids expressing TRIM5α and its fragments fused with GFP tag were constructed successfully. The TRIM5α and its fragments from human, rhesus monkey, and AGM were all located in the cytoplasm, among which the hTRIM5α, rhTRIM5α, AGMTRIM5α, rh-TRIM5α CC/B30.2, AGM-TRIM5α CC/B30.2 were distributed as oligomer. But h-TRIM5α CC/B30.2 and TRIM5α B30.2 from different species were distributed evenly in the cytoplasm. The inhibition rate of pseudotyped virus infection was about 70% for rh-TRIM5α, AGM-TRIM5α, rh-CC/B30.2 and AGM-CC/B30.2, and that was less than 20% for h-TRIM5α and different species B30.2.
Conclusion TRIM5α CC/B30.2 domain of rhesus monkey and AGM could inhibit HIV-1. Reliable experimental data helping determine the TRIM5α functional domains were obtained.

Key words: TRIM5α, HIV-1, virus