TABLE 1.
Oligonucleotides used in charged-to-alanine scanning mutagenesis of Ty3 IN
| Ty3 IN mutanta | Mutation(s) | Oligonucleotide | Sequence |
|---|---|---|---|
| 11A(2) | D11A, E13A | 364 | 5′GATTTCCAGCTTGCTGTGGCGATAGGTCG3′ |
| 20A(2) | K20A, D22A | 365 | 5′CATAATGGGGCTGATGCGTAGTAAG3′ |
| 33A(2) | K33A, E34A | 366 | 5′GTTGTGTCAATGCTGCCATATGAATTAAG3′ |
| 43A(2) | E43A, D44A | 367 | 5′GGCTGACATAGCTGCAGGTGTGAC3′ |
| 53A(2) | K53A, K54A | 368 | 5′GATAGTTCGAGTGCCGCCTGGTAAC3′ |
| 62A(2) | R62A, K63A | 369 | 5′GGGAATAATTCGCTGCGAAGGTCTC3′ |
| 69A(2) | D69A, E70A | 370 | 5′GTAATAGATCATTGCGGCTTCTAGGG3′ |
| 76A(2) | D76A, R77A | 371 | 5′GGTACTACTAGTGCGGCTTGGTAATAG3′ |
| 412A(2) | E412A, D414A | 373 | 5′GAGAGTTTAAAGCTAGTGCGTAGGCGTTATC3′ |
| 419A(3) | K419A, K420A, K421A | 374 | 5′CTCTGTGCGCTGCCGCGTGAGAG3′ |
| 431A(2) | K431A, K432A | 375 | 5′GGTATACAAACGCTGCCAGGAATTG3′ |
| 436A(2) | R436A, D438A | 376 | 5′GGGTACGCGGCTGGAGCGTATACAAAC3′ |
| 442A(2) | K442A, K444A | 377 | 5′GCTGATTGGTGCATTCGCTGGGTACGC3′ |
| 450A(2) | E450A, R451A | 378 | 5′CTCTCTTAATTGCTGCAGTGGAGCTG3′ |
| 453A(2) | K453A, R454A | 379 | 5′CTTCGTGTGCTGCCGCAATTCTTTC3′ |
| 477A(2) | D477A, D479A | 380 | 5′GAAAGTGTTGGGGCTACAGCTTGCATGTG3′ |
| 488A(2) | E488A, E490A | 381 | 5′GGCAAAATGCAGCTGCTGAGTATTC3′ |
| 496A(2) | E496A, R497A | 382 | 5′GATCTTCGTGTTGCTGCGGGAATTTGG3′ |
| 499A(2) | R499A, R500A | 383 | 5′GCTAATATTGATGCTGCTGTTCTTTCG3′ |
| 519A(1) | E519A | 384 | 5′CAACATCTTCCGCTCTCTCAGGG3′ |
a
Ty3 IN mutants are referred to by the codon position of the first amino acid residue in the mutagenized cluster, followed by an A to indicate a change to alanine and a number in parentheses to indicate how many residues were changed in the respective cluster.