Table 2.

Nucleotide sequence of PCR primers used to introduce mutations

Primer name	Sequence 5′ → 3′	Nucleotide positionsa
ME46−3′b	GTTATTGGTCTTAAAGGTACCTGAGGTCTGACTG	9035−9002
ME46−5′c	CAATATCGAGATCTTCAGACCTGGTGGAGGAGAT	7610−7646
ME46−770Ed	CTCTTCAGCTACCACGAATTGAGAGACTTACTC	8517−8549
ME46−772E	AGCTACCACCGCTTGGAAGACTTACTCTTGATTG	8523−8556
ME46−788E	GAACTTCTGGGACGCGAGGGGTGGGAAATCCTC	8571−8603
89.6−3′e	CAGACGGGCACACACTACTTGAAGCACTCA	9655−9626
89.6−5′f	GGCACTGAAGGAAATGACATAATCACACTC	7443−7472
89.6−770E	CTCTTCCTCTACCACGAATTGAGAAACTTACTC	8517−8549
89.6−772E	TCTTCCTCTACCACCTCTTGGAAAACTTACTCTTGATTGTA	8518−8558
89.6−788E	GAACTTCTGGGACGCGAGGGGTGGGAAGCCCTCAAATATTGG	8571−8612
89.6−846E	CTATTCGCAACATACCTACAGAGATCAGACAGGGCTTGGAAAG	8740−8782
89.6−848E	GCAACATACCTACAGAGATCGAGCAGGGCTTGGAAAGGGCTTT	8746−8788

^aRelavant segments of the HIV-1 ME46 and HIV-1 89.6 env gene DNA sequences were entered into the HIV/SIV Sequence Locator Tool at the Los Alamos Sequence Database to give the nucleotide positions relative to HXB2.

^bME46−3′ and ME46−5′ represent the non-mutagenic primers used in the overlap PCR mutagenesis protocol.

^cME46−3′ and ME46−5′ represent the non-mutagenic primers used in the overlap PCR mutagenesis protocol.

^dThis table lists the sense strand mutagenic primer to pair with the 3′ non-mutagenic primer for the first step of the overlap PCR mutagenesis in one reaction. An appropriate corresponding antisense primer to pair with the 5′ non-mutagenic primer was also used (sequence not shown) in a parallel reaction. The sequences in bold represent the codon changed to introduce the glutamic acid residue mutation.

^e89.6−3′ and 89.6−5′ represent the non-mutagenic primers used in the overlap PCR mutagenesis protocol.

^f89.6−3′ and 89.6−5′ represent the non-mutagenic primers used in the overlap PCR mutagenesis protocol.