TABLE 1.
Mutant | Oligonucleotide | Oligonucleotide sequence (5′ → 3′)a |
---|---|---|
E-TRS-N | N-800 AscI VS | TTGGCGCGCCACTCCTTGCTTGAACTAAACAAAATG |
3′-AscI RS | TTGGCGCGCCTTAGTTCGTTACCTCATCAATTATC | |
TRS-N | N-120 AscI VS | TTGGCGCGCCTTGAAAGCAAGTAGTGCGACTGG |
3′-AscI RS | ||
E-TRS-GFP | E-800 AvrII VS | TTCCTAGGACTCCTTGCTTGAACTAAACAAAATG |
GFP 5′ RS | CAGCTCCTCGCCCTTGCTCACCATTTAGAAGTTTAGTTA | |
GFP 5′ VS | ACATATGGTATAACTAAACTTCTAAATGGTGAGCAAGGGC | |
3′AscI GFP RS | AAGGCGCGCCGGAAGAAAGCGAAAGGAGCGGGCGC | |
E-TRS3a-N | E-800 AvrII VS | |
E-120 AvrII RS | AACCTAGGCTTCTTGCCAACAAGTGTGTAGACAATAGTC | |
E1-TRS-N | E-800 AvrII VS | |
E-460 AvrII RS | AACCTAGGGACACTTGGTATTCCGAGTATGCATTAAAAATC | |
E2-TRS-N | E-740 AvrII VS | TTCCTAGGTGGAACTTCAGCTGGTCTATAATATTGATC |
E-160 AvrII RS | AACCTAGGCATAGCTTCTTCCTAATGCACTAACGCAAAG | |
E3-TRS-N | E-460 AvrII VS | TTCCTAGGCAGATATGTAATGTTCGGCTTTAGTATTGC |
E-120 AvrII RS | ||
M1-TRS-N | Rep Mut 2.1 VS | GTCCAGATTACATATGTTCGGCTTTAGTAT |
Rep Mut 2.1 RS | CCGAACATATGTAATCTGGACACTTGGTAT | |
M2-TRS-N | Rep Mut 2.2 VS | AAAATTATATGTAATGGTATAACTAAACTT |
Rep Mut 2.2 RS | TATACCATTACATATAATTTTTCTTGCTCA | |
M4-TRS-N | Rep Mut 2 A1G VS | GTGTCCAGGTATGTAATGTTCGGCTTTAGTA |
Rep Mut 2 A1G RS | AGCCGAACATTACATACCTGGACACTTGGTATTC | |
M5-TRS-N | Rep Mut 2 A1C VS | GTGTCCAGCTATGTAATGTTCGGCTTTAGTA |
Rep Mut 2 A1C RS | AGCCGAACATTACATAGCTGGACACTTGGTATTC | |
M6-TRS-N | Rep Mut 2 A1C-U9C VS | GTGTCCAGCTATGTAACGTTCGGCTTTAGTA |
Rep Mut 2 A1C-U9C RS | AGCCGAACGTTACATAGCTGGACACTTGGTATTC | |
M7-TRS-N | Rep Mut 2 CC-U9C VS | GTGTCCCCATACGTAATGTTCGGCTTTAGTA |
Rep Mut 2 CC-U9C RS | AGCCGAACATTACGTATGGGGACACTTGGTATTC |
The mutated nucleotides are shown in boldface. The restriction endonuclease sites used for cloning are underlined (AvrII, CCTAGG; AscI, GGCGCGCC).