Table 1 Sequences of oligonucleotide primers used in this study
Target gene | Gene position on SF301 genome or virulent plasmid pCP301 | Primer* | Primer sequence (5′→3′) | Length (bp) |
|---|---|---|---|---|
Primers for detection of virulence-associated genes of S. flexneri by mPCR | ||||
ipaH | 1422225–1422835 ** | ipaH-F | CCTTGACCGCCTTTCCGATA | 611 |
| Â | Â | ipaH-R | CAGCCACCCTCTGAGAGTACT | Â |
ial | 133550–133869*** | ial-F | CTGGATGGTATGGTGAGG | 320 |
| Â | Â | ial-R | CCAGGCCAACAATTATTTCC | Â |
set1B | 3069523–3069669** | set1B-F | GTGAACCTGCTGCCGATATC | 147 |
| Â | Â | set1B-R | ATTTGTGGATAAAAATGACG | Â |
Primers for amplifying int , orf30 , sigA and pic on PAI-1 of S. flexneri 2a | ||||
int | 3052736–3053998** | int-F | ATGGCACTGACTGACGCAAA | 400 |
| Â | Â | int-R | TGCCGATAAAGGGGAAAACG | Â |
orf30 | 3096187–3097975** | orf30-F | CTTATCACTGAGCGTCTGGT | 1,102 |
| Â | Â | orf30-R | GTGAAATTCCTGCCTCAATA | Â |
sigA | 3060437–3064294** | sigA-F | AGTCATATTACAGGTGGATTAG | 1,866 |
| Â | Â | sigA-R | TATACTCAGGGTTGCGTTTT | Â |
pic | 3067737–3070949** | pic-F | AGAACATATACCGGAAATTC | 1,219 |
| Â | Â | pic-R | ACCCTGACGGTGAATAAACT | Â |
Primers for homologous recombination to construct pic knockout strain | ||||
upstream of pic | 3067236–3067745** | uppic-F-NotI | AAGCGGCCGCCATAGCAGACTGGCCGGTCAACC | 520 |
| Â | Â | uppic-R-XbaI | CCTCTAGAATGTTCTGATGTGGGGGTAAAGGGC | Â |
downstream of pic | 3071850–3072358 ** | downpic-F-XbaI | CCTCTAGAATTCACTATGGATTCTCCATGAT | 517 |
| Â | Â | downpic-R-BamHI | AAGGATCCCGTCGTCCGTCTGGCACC | Â |
upstreamof pic | 3066436–3072733** | Upuppic-F | GCTGAACTGC TGGAGCCGCT | 1176 |
downstream of pic | Â | Downdown Pic-R | CAGCGGCGAAATACTGTACC | Â |
pic coding frame work | 3067737–3070949** | pic-pSC-F-PfMlI | AAACCATCGAATGGATGCAGGACGATTTCGATGCCCCCGTAGAC | 3,213 |
| Â | Â | pic-pSC-R-AclI | TTTAACGTTTCAGAACATATACCGGAAATTCGCGTT | Â |