CONSTRUCTION OF GAG, POL, AND ENV SPECIFIC RIBOPROBES FOR CONFIRMATION OF HIV-1 SPECIFIC POLYMERASE CHAIN-REACTION PRODUCTS (VOL 48, PG 167, 1994)

Conserved regions of the gag, pol, and env genes of HIV-1 pBH10 DNA (g ag nucleotides(nt)1508-1652, pol nt 2811-3118, env nt 7792-7934; Ratne r et al., 1985) were amplified by the polymerase chain reaction (PCR) using oligonucleotides complementary to the termini of these regions a s primers. Primer areas of the amplified DNA were then removed by dige stion with restriction endonucleases, and the internal fragments purif ied and cloned in both orientations into the 'riboprobe' transcription vector pGEM-5Z. Riboprobes made from these plasmids did detect the sp ecific sequences of pBH10 DNA and of HIV-1 DNA amplified by PCR from c linical material. The riboprobes will be useful to confirm the specifi city of PCR-amplified fragments of lymphocyte DNA obtained from infant s of HIV-infected mothers and from high risk, but seronegative contact s of HIV-1 infected individuals.