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. 2010 Dec 21;10:324. doi: 10.1186/1471-2180-10-324

Table 3.

Primers

Name Nucleotide sequence (5' to 3')a
Construction of the deletion mutants
relA-FW CGCCATCCCGCAATGGTTTACATAA
relA-RV TCATTGTTCTGGCCATAACAGC
spoT-FW CTTGAAAACCATCATTCGCGCTGAACG
spoT-RV TCTGCGGTACGAATGATTGCAGAAACG
stm3169-red-FW ACGTTCATTCACAACATCAGCGGTATTACTGGCCGGCTGTGTGTAGGCTGGAGCTGCTTC
stm3169-red-RW ACATATTCTCGATGTATTCCAGATCCTTCGCTGACTGAGCCATATGAATATCCTCCTTAG
sseF-red-FW AACAGAACGAAATATGAAAATTCATATTCCGTCAGCGGCAGTGTAGGCTGGAGCTGCTTC
sseF-red-RW TGTCCATTAATGCAGGTGTAGTAGCAGATTGACAGAGCGCCATATGAATATCCTCCTTAG
pAC-tet-FW TTGGTAGCTCAGAGAACCTTCGAAAAACCG
pAC-tet-RV TCGCTCGCGTATCGGTGATTCATTCGCTA
Construction of plasmids for the complementations
relA-FW2 AGGCTCGAGGTCGCGGTAAGAAGT
relA-RV2 ACAAGCTTACTGTCTGGGGTTTAC
ssrB-FW GGGCTCGAGGAATATAAGATCTTATTAGTA
ssrB-RV CCCGGATCCATACTCTATTAACCTCATTCT
stm3169-FW CCGCTCGAGAACACACGTTCATTCACAACATCAG
stm3169-RV GGAAGATCTATTCTCGATGTATTCCAGATCCTTC
Construcion of the lacZ fusions
ssaG-Pro-FW AAAGTCGACCAAATGCTCAGGTAGGAGGGC
ssaG-Pro-RV AAAGGATCCATCATCGATTCTGGGTTGAGC
stm3169-Pro-FW ACGCGTCGACGACGATTTAGCCGGTATGAAAATCA
stm3169-Pro-RV CGGGATCCTTACATATTCTCGATGTATTCCAGA
Comfirmation of gene expression by qRT-PCR
gyrA-FW AAGAGCTCCTATCTGGATTATGC
gyrA-RV TATTTACCGATTACGTCACCAAC
relA-FW ATTGTGCCATTCACCTATCAGTT
relA-RV GATATTTTTGTCACGATCCTGCT
invF-FW ATCGCTGCTGAATAGTGTAGAAG
invF-RV CATTTGTCTGCCAATTGAATAAT
stm0209-FW CCTGAACGTAGAAAATTACGAGA
stm0209-RV GTCAGGTTTTTCACCATGTTACT
stm0435-FW GTCAATCAGTTGCTCGATATTCTG
stm0435-RV TTTAATCAGCTTGACGATTTTCTTC
stm0748-FW TGAACCTGTACGTTATGGATCTC
stm0748-RV CGCCGTTAATGTTCATTTTATAC
stm0781-FW GAAGGCAAGATCACCGTATTT
stm0781-RV CTGATCAGCAGAGATGAAGAGAT
stm1478-FW ACAAAAGTTGAGGAGCTGAATAAAG
stm1478-RV GCCACTGACGCGTAATATGATAA
stm1720-FW TTGGTTGTAAAATTGAAGACAAAGG
stm1720-RV GTCCCCTTCAGGATAAAGGTGTAAT
stm1746-FW CGAATTATTCCAGAAACTGAAGAAA
stm1746-RV ATCGCCCTGATTTTTAACCTTATTA
stm2638-FW TATTCTGACGGTCTGTTTAGCTTTT
stm2638-RV GTACTGCCCTGAATTTGATACTGTC
stm3169-FW GTTACCAGAATAATGTCGCAGCTAT
stm3169-RV AATCATCCACATAAAAAGAATCTGG
stm3318-FW CAAACTCAGCCTTAATCTTATGC
stm3318-RV ACTTTATCGGCGTTGATCTTAAT
stm4319-FW ATTAGTATTATCCGAGGCCAGAC
stm4319-RV CAGTCTTGCAAACTCTACTGCTC
stm4403-FW ATTGATATTCACAGCAACAAACC
stm4403-RV AGGTCAGGTTTTTAATACGTTCC
stm4405-FW CGAACTGACGTTGAATAAAGATGAG
stm4405-RV AATTGTGGTCGTCTGGTATCTGT

aUnderlined part indicates P1 or P2 site for pKD4, or restriction sites.