Table 2.
Primers used in this study
| Primer namea | Sequence (5′→3′)b |
Use | |
|---|---|---|---|
| Forward | Reverse | ||
| EF3002 (slyA) | GTGCGCGAAAATCAACACT | CATTTTTACGCATCCGGACTA | RT-PCR |
| EF3002R1 | CCTTAAATTCAATATTACTG | 5′RACE-PCR | |
| EF3002R2 | CTTTTCTTGGATAATCCCAGG | 5′RACE-PCR | |
| EF3002U | CCGGTCGACATGAGTTTCTAGAGGAGTTCCTA | CGCGGATCCGGCCCGCGCAATCATGCCAATTTCTC | Cloning in pMad, RT-PCR |
| EF3002D | CGCGGATCCGTCAGCGAAGATTGGGAATTTGT | GAAGATCTCGATTATTTGTTCACTTTGAACG | Cloning in pMad, RT-PCR |
| EF3002int | GAGAAATTGGCATGATTGCGCGGGCC | TCTGTGGCATAAATCCGCTTA | RT-PCR |
| EF3001int | TAAATCGTGAGACGGCACAA | AAAGGAAAGAAGCTGGGGAC | RT-PCR |
| EF3002tot | GACGCTATAACTAATTTTATCTTAC | TTAATAATTTCGAGTGTTCCC | Cloning in pCU1 |
| PU/PR | GTAAAACGACGGCCAGT | CAGGAAACAGCTATGAC | Cloning verification |
a
From the annotated sequence available at http://www.tigr.org. EF3002U and EF3002D are primers used to clone upstream (U) and downstream (D) sequences of slyA in order to construct the deletion mutant by a double-crossover, and EF3002tot is for the in trans-complemented strain.
b
The underlined sequences are restriction enzyme recognition sites.