TABLE 1.
Oligonucleotides used in the course of the study
| Primer function and name | Sequence (5′ to 3′)a |
|---|---|
| PCR | |
| BsCRASP-3 145(+) BamHI | GCTGTTTTTGCACTGTTTGGATCCTGTGGAAATTTTAC |
| BsCRASP-3 3nc(−) XhoI | ATTCATAATTATTCTCTTCTCGAGTTTGAATTTCTA |
| TISAR32 | TTCATAATTATTCTCTTCTCGAGTTTG |
| TISAR33 | CACCTGTGGAAATTTTACAAGTAGTTTA |
| pGEX(+) | GGGCTGGCAAGCCACGTTTGGTG |
| pGEX(−) | CCGGGAGCTGCATGTGTCAGAGG |
| Site-directed mutagenesis | |
| BsCRASP-3 E77G(+) | CGGGTTTAAATGCTACTGGGGGGCATTCAGCTACGTTATTTTC |
| BsCRASP-3 E77G(−) | GAAAATAACGTAGCTGAATGCCCCCCAGTAGCATTTAAACCCG |
| BsCRASP-3 H79R(+) | GTTTAAATGCTACTGAGGGGCGTTCAGCTACGTTATTTTCATTAG |
| BsCRASP-3 H79R(−) | CTAATGAAAATAACGTAGCTGAACGCCCCTCAGTAGCATTTAAAC |
| BsCRASP-3 E131K(+) | GAGATAATAACAAAGATGAAAACCATTGATGGTTCTGAAC |
| BsCRASP-3 E131K(−) | GTTCAGAACCATCAATGGTTTTCGTCTTTGTTATTATCTC |
| BsCRASP-3 Q115E(+) | GTTTATATTATGGATATAGGGAAGAGAGCACTGAAAAGGGTATCC |
| BsCRASP-3 Q115E(−) | GGATACCCTTTTCAGTGCTCTCTTGCCTATATCCATAATATAAAC |
a
Underlining indicates introduced substitution(s) of the respective residue(s).