TABLE 1.

Oligonucleotides used for site-directed mutagenesis

MutantOligonucleotideOligonucleotide sequence (5′ → 3′)a
E-TRS-NN-800 AscI VSTTGGCGCGCCACTCCTTGCTTGAACTAAACAAAATG
3′-AscI RSTTGGCGCGCCTTAGTTCGTTACCTCATCAATTATC
TRS-NN-120 AscI VSTTGGCGCGCCTTGAAAGCAAGTAGTGCGACTGG
3′-AscI RS
E-TRS-GFPE-800 AvrII VSTTCCTAGGACTCCTTGCTTGAACTAAACAAAATG
GFP 5′ RSCAGCTCCTCGCCCTTGCTCACCATTTAGAAGTTTAGTTA
GFP 5′ VSACATATGGTATAACTAAACTTCTAAATGGTGAGCAAGGGC
3′AscI GFP RSAAGGCGCGCCGGAAGAAAGCGAAAGGAGCGGGCGC
E-TRS3a-NE-800 AvrII VS
E-120 AvrII RSAACCTAGGCTTCTTGCCAACAAGTGTGTAGACAATAGTC
E1-TRS-NE-800 AvrII VS
E-460 AvrII RSAACCTAGGGACACTTGGTATTCCGAGTATGCATTAAAAATC
E2-TRS-NE-740 AvrII VSTTCCTAGGTGGAACTTCAGCTGGTCTATAATATTGATC
E-160 AvrII RSAACCTAGGCATAGCTTCTTCCTAATGCACTAACGCAAAG
E3-TRS-NE-460 AvrII VSTTCCTAGGCAGATATGTAATGTTCGGCTTTAGTATTGC
E-120 AvrII RS
M1-TRS-NRep Mut 2.1 VSGTCCAGATTACATATGTTCGGCTTTAGTAT
Rep Mut 2.1 RSCCGAACATATGTAATCTGGACACTTGGTAT
M2-TRS-NRep Mut 2.2 VSAAAATTATATGTAATGGTATAACTAAACTT
Rep Mut 2.2 RSTATACCATTACATATAATTTTTCTTGCTCA
M4-TRS-NRep Mut 2 A1G VSGTGTCCAGGTATGTAATGTTCGGCTTTAGTA
Rep Mut 2 A1G RSAGCCGAACATTACATACCTGGACACTTGGTATTC
M5-TRS-NRep Mut 2 A1C VSGTGTCCAGCTATGTAATGTTCGGCTTTAGTA
Rep Mut 2 A1C RSAGCCGAACATTACATAGCTGGACACTTGGTATTC
M6-TRS-NRep Mut 2 A1C-U9C VSGTGTCCAGCTATGTAACGTTCGGCTTTAGTA
Rep Mut 2 A1C-U9C RSAGCCGAACGTTACATAGCTGGACACTTGGTATTC
M7-TRS-NRep Mut 2 CC-U9C VSGTGTCCCCATACGTAATGTTCGGCTTTAGTA
Rep Mut 2 CC-U9C RSAGCCGAACATTACGTATGGGGACACTTGGTATTC
  • a The mutated nucleotides are shown in boldface. The restriction endonuclease sites used for cloning are underlined (AvrII, CCTAGG; AscI, GGCGCGCC).