Table 2 Primers used in this study
From: The global regulators ArcA and CytR collaboratively modulate Vibrio cholerae motility
Name | Sequence (5′–3′) | Amplifified fragment |
|---|---|---|
Primers for construction of mutants | ||
∆arcA-S-F | GCTCTAGACGATCAAGCATTGCTGTAAA | ∆arcA-S (500) |
∆arcA-S-R | AAAGAAGAGGTAGCGTTACCTAAACTTGTGA | |
∆arcA-X-F | GGTAACGCTACCTCTTCTTTTATATCTAATTAG | ∆arcA-X (500) |
∆arcA-X-R | CGGAGCTCAACATCATGCCGGTGAGAG | |
∆arcA-SX (1000) | ||
arcA-F | ATGCAAACCCCGCAGATCCTT | arcA (717) |
arcA-R | TTAATCTTCTAAATCACCACAG | |
∆cytR-S-F | CGGGGTACCTCCGAGGACGACACGATAC | ∆ cytR-S (512) |
∆ cytR-S-R | GTAAAAATACCCCACCTTCGAAACCGA | |
∆ cytR-X-F | CGAAGGTGGGGTATTTTTACCCTCTTTTCTCTATCG | ∆ cytR-X (534) |
∆ cytR-X-R | CGAGCTCTTTCGAGCTGAAGCCAATC | |
∆ cytR-SX (1046) | ||
cytR-F | ATGGCGACAATGAAGGATGT | cytR (1015) |
cytR-R | AGGTGGGTTACTTCTTGCTTG | |
Primers for identification of plasmid | ||
pRE112-U-F | CACTGTTCGTCCATTTCCG | pRE112-UD (567) |
pRE112-D-R | TTCGTCTCAGCCAATCCCT | |
pRE112-U-arcA-D (1284) | ||
pRE112-U-cytR-D (1582) | ||
pBAD33-U-F | AACAAAGCGGGACCAAAG | pBAD33-UD (529) |
pBAD33-D-R | AGAGCGTTCACCGACAAA7 | |
pBAD33-U-arcA-D (1246) | ||
pBAD33-U-cytR-D (1544) | ||
pET28a-U-F | TAATACGACTCACTATAGGG | pET28a-UD (318) |
pET28a-D-R | GCTAGTTATTGCTCAGCGG | |
pET28a-U-arcA-D (1035) | ||
pET28a-U-cytR-D (1395) | ||
pET28a-U-crp-D (918) | ||
Primers for construction of complemented strain | ||
∆arcA::ParcA-F | CGAGCTCTAAATCAACAAAGTGATTGGAA | ∆arcA::ParcA (732) |
∆arcA::ParcA-R | GGGGTACCTTAATCTTCTAAATCACCACAG | |
∆cytR::PcytR-F | GGGGTACCATGGCGACAATGAAGGATGTTG | ∆cytR::PcytR (1031) |
∆ cytR::P cytR -R | GCTCTAGAGCTTACTTCTTGCTTGGCGGCG | |
Primers for protein cloning | ||
pET28a-arcA-F | CGGGATCCATGCAAACCCCGCAGATCCT | pET28a-arcA (734) |
pET28a-arcA-R | CCGCTCGAGTTAATCTTCTAAATCACCAC | |
pET28a-cytR-F | CGCGGATCCATGGCGACAATGAAGGATG | pET28a-cytR (1095) |
pET28a-cytR-R | CCGCTCGAGTTACTTCTTGCTTGGCGG | |
pET28a-crp-F | CGCGGATCCATGGTTCTAGGTAAACCTCA | pET28a-crp (617) |
pET28a-crp-R | CCGCTCGAGTTAGCGAGTGCCGTAAACCA | |
Primers for bioluminescent reporter assays | ||
pMS402-cytR-F | CGGGATCCCTTTTACTACAAACGCCGAAT | pMS402-cytR (1032) |
pMS402-cytR-R | CCGCTCGAGGTATTTTTACCCTCTTTTCTCTATC | |
pMS402-flrA-F | CGGGATCCAACGTTTAGGTAAAGCCTTGG | pMS402- flrA (1484) |
pMS402-flrA-R | CCGCTCGAGAGGTGAGATTATTTGCCTTTATTAT | |
pMS402-fliK-F | CGGGATCCACTGCGTCAAATTGAACAGTACC | pMS402-fliK (2042) |
pMS402-fliK-R | CCGCTCGAGAGTGGAATTGAAGTCTGAGCATG | |
Primers for qRT-PCR | ||
16S rRNA-F | GTGTACGGTGAAATGCGTAGAG | 275 bp |
16S rRNA-R | GCGTGGACTACCAGGGTATCTAAT | |
qRT- cytR -F | ATTCGCGGTATTGAAGATGC | 189 bp |
qRT- cytR -R | AGGCGGTAGGTTTTTCTGCT | |
qRT- flrA -F | CCTGAAGGGGTGAATCTCAA | 157 bp |
qRT- flrA -R | GCATGTTGTATTTGCGCATC | |
qRT- fliK -F | CTCAAACCGTAGCGGTCAAT | 235 bp |
qRT- fliK -R | TGTACCAGTTGCGACTCAGC | |
Primers for EMSA | ||
EMSA-cytR-F | ATCGCGTTTTATAACGCTGAT | 200 bp |
EMSA-cytR-R | CTAGAAATCATGGCCATAACCA | |
EMSA- flrA -F | ATAAAGTCAGCTTGGGATCAAA | 300 bp |
EMSA- flrA -R | AGGTGAGATTATTTGCCTTTATTAT | |
EMSA- fliK -F | GTCAAAAACGGAAATCCTATCA | 300 bp |
EMSA- fliK -R | AGTGGAATTGAAGTCTGAGCAT | |