TABLE 1.
Primers used to create new CA mutations
| HIV CA mutant | 5′-3′ (forward) sequence (mutated codons underlined) | Altered restriction site |
|---|---|---|
| Q4A | TACCCCTATAGTCGCGAACCTCCAGGG | NruI |
| Q13A | GGCAAATGGTGCATGCGGCCATATCACCTAG | SphI |
| R18A/N21A | CATATCACCTGCAACTTTAGCTGCATGGGTA | NsiI destroyed |
| P38A | CCAGAAGTAATCGCGATGTTTTCAGC | NruI |
| E45A | CATGTTTTCAGCCTTATCGGCAGGAGCCACC | BglI |
| T54A | CACAAGATTTGAATGCCATGCTAAAC | BsmI |
| T54A/N57A | AGATTTAATGCCATGCTAGCCACAGTGGG | NheI |
| Q63A/Q67A | GGGGGGACATGCAGCAGCCATGGCAATGTTAAAAG | DsaI |
| K70A | CCATGCAAATGCTAGCAGAGACCATC | NheI |
| E71A | CATGCAAATGCTTAAGGCGACCATCAAT | AflII |
| N74A | GTTAAAAGAGACGATCGCTGAGGAAGCTG | PvuI |
| E75A/E76A | GACCATCAATGCGGCGGCCGCAGAATGGG | NotI |
| A78D/E79A/R82A | GAGGAAGCTGACGCATGGGATGCATTGCATCCAG | PstI destroyed |
| R100A/S102A | GAGAGAACCAGCGGGAGCGGACATAGCAGG | BsrBI |
| T107A/T108A | CATAGCAGGAGCTGCTAGTACCCTTC | SpeI destroyed |
| T110A/Q112A | GAACTACTAGCGCCCTTGCGGAACAAATAG | SpeI destroyed |
| G116A | GAACAAATAGCATGGATGACA | None |
| T119A | CAAATAGGATGGATGGCACATAATCC | None |
| E128A/R132Aa | CCCAGTAGGAGCAATCTATAAAGCTTGGATAATCC | HindIII |
| L136D | TAAAAGATGGATAATCGATGGATTAAATAAAATAG | ClaI |
| N139A | GATAATCCTGGGACTCGCGAAAATAGTAAG | NruI |
| R143A | AATAAAATAGTGGCCATGTATAGCCC | MscI |
| D152A | TACCAGCATTTTAGCCATAAGACAAG | BsmI destroyed |
| K158A | ACAAGGACCAGCTGAACCCTTTAG | PvuII |
| K158D | ACAAGGACCAGATGAACCCTTTA | None |
| K158R | GACAAGGACCTCGAGAACCCTTTA | XhoI |
| K158Q | ACAAGGACCACAAGAACCCTTA | None |
| D163A | CCCTTTAGAGCCTACGTAGACCGA | SnaBI |
| K170A | CCGATTCTATGCGACGCTAAGAGCC | HgaI |
| Q176A | AAGAGCCGAGGCAGCTTCACAAG | HindIII destroyed |
| E180A | CGAGCAAGCTACACAAGCGGTAAAAAAT | HindIII destroyed |
| W184A | AGGTAAAAAATGCGATGACAGAAACGTTGTTGGTCCAA | Psp 1406I |
| M185A | AGGTAAAAAATTGGGCGACAGAAACGTTGTTGGTCCAA | Psp 1406I |
| Q192A | CTTGTTGGTCGCGAATGCGAACC | NruI |
| D197A | TGCGAACCCAGCATGCAAGACTATTTT | SphI |
| D197N | TGCGAACCCAAACTGTAAGACTAT | XhoI |
| D197E | CAAAATGCGAATCCGGAGTGGAAGACTATTTTAAAAGC | BbsI |
| K203A | GATTGTAAGACAATATTAGCAGCATTGGGA | SspI |
| P207A | AAGCATTGGGCGCCGGAGCGACAC | EheI |
| E212A | CAGGAGCGACGCTAGCAGAAATGATGAC | NheI |
| Q219A | TGACAGCATGCGCAGGAGTGGGGG | FspI |
| P224A | GAGTGGGGGGAGCGGGCCATAAAG | BsrBI |
a
R132A was also inadvertently made as a single mutant, but it was fully infectious and showed a phenotype similar to that of the double mutant, so it is not listed separately.