Table 1 Strains and plasmids used in this study
Strains and plasmids | Genotype or description | Source or references |
|---|---|---|
Strains | ||
Kp4 | Mutant from Klebsiella pneumoniae ATCC10280 by UV | Lab collection |
E. coli DH5α | Cloning strain | Wei et al. (2012) |
E. coli BL21 | Cloning strain | Lab collection |
JJQ01 | Kp4 ΔldhA::apr | This study |
JJQ02 | Kp4 ΔldhAΔdhaT::apr | This study |
Plasmids | ||
pUC18-kan-aldH | pUC18-kan carries aldH from E. coli | |
pIJ773 | Apramycin resistance, carries disruption cassette aac(3)IV | Wei et al. (2012) |
pMD18-T-2028/a | Ampicillin resistance, carries sequences ldhA::FRT::apr | Wei et al. (2012) |
pMD18-T simple | Cloning vector | Sangon Biotech Co., Ltd. |
pDK6-Red | Carries red recombinase gene | Wei et al. (2012) |
pDK6-Flp | Carries Flp recombinase gene | Wei et al. (2012) |
pMD18-T-dhaT | Ampicillin resistance, carries gene dhaT | This study |
pMD18-T-FRT-aac(3)IV | Ampicillin resistance, carries disruption cassette aac(3)IV | This study |
Primers | Sequences (from 5′ to 3′) |
|---|---|
ldh-s1 | AGAGCGCACAGGACCACTATCCA |
ldh-a1 | TCGGCGAGCTTATAGACCAGCGT |
dhaT-T-F | CCATTACCGGCAATAGCGGAACT |
dhaT-T-R | TCTCGGCCATATTGCGGTCGATA |
dhaT-FRT-F1 | GCATTATAACCTGAAGCGAGAAGGTATATTATTCCGGGGATCCGTCGACC |
dhaT-FRT-R1 | AAAGCGACATGACGGCCCCCTCGTTAACACTGTAGGCTGGAGCTGCTTC |
FRT-apr-F | ATTCCGGGGATCCGTCGACC |
FRT-apr-R | TGTAGGCTGGAGCTGCTTC |
T-F | AATATACCTTCTCGCTTCAGGTTATAATGC |
T-R | GTGTTAACGAGGGGGCCGTCATGTCGCTTT |