FIG. 2.

DNA sequences of 30 protease recombinants of the HIV-1 pNG-PRL library. WT, wild type. Thirty randomly selected clones were isolated and sequenced. Conserved nucleotides are indicated by a dot. For DNA sequencing, InstaGene (Bio-Rad Laboratories) was used for preparation of DNA from HIV-1-infected MT-2 cells. A 706-bp fragment containing a viral protease-encoding sequence was amplified by PCR with primers PR5 (5′-GTT AAG TGT TTC AAT TGT GGC AAA GAA GGG C-3′) and PR12 (5′-CTC TGT ACA AAT TTC TAC TAA TGC-3′). The PCR products were purified via 1% agarose electrophoresis and cloned into StuI-digested pCRΔZ vector created from pCR-blunt (Invitrogen) by deletion of a PmlI fragment containing a Zeocin cassette. Cloned sequences were sequenced with ABI Prism 377 (Applied Biosystems, Inc., Foster City, Calif.).