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TABLE 3.

Evaluation of the nef open reading frame in animals inoculated with Tyr-721 mutantsa

Virus Animal Time (wk) Total no. of clones No. of clones with following amino acid encoded by nef codon 92:
Stop (TAA) Glu (GAA) Ser (TCA) Tyr (TAC) Gln (CAA) Lys (AAA) Leu (TTA) Glu (GAG) Asp (GAC)
SIVmac239 N2V 8 8 1 1 2 3 1
K7K 8 9 3 1 2 1 2
N7K 4 7 4 1 2
SIVmac239Y/I VF3 2 11 6 3 2
N8G 4 10 6 1 3
N8G 12 6 1 3 2
N8G 44 6 5 1
N8G 56 7 3 2 1 1
N8G 80 10 3 3 4
SIVmac239ΔGY WD1 2 11 7 2 2
WD1 44 11 9 1 1
WD1 56 2 1 1
AGP 4 9 9
AGP 8 10 3 3 4
AGP 25 5 2 3
AGP 52 5 1 4
2AJ 2 11 10 1
a

DNA from peripheral blood lymphocytes of the animals indicated was amplified using nested primers that flanked the premature termination codon in nef at position 92 that was present in all viruses during the initial infection. Fragments were cloned into pCDNA and sequenced. Codon 92 and the predicted amino acids are shown at the indicated weeks postinoculation. nef reverted to an open reading frame in wild-type SIVmac239 and the Y/I and ΔGY mutants, although persistence of the stop codon was seen in some clones for animal N8G at 56 and 80 weeks.