TABLE 1.

Oligonucleotides used for PCR amplification, site-directed mutagenesis, and chimeric RpoB construction

Oligonucleotide name and purpose	Sequence (5′→3′)a	Tm (°C)
Primers for RpoB and RpoC subunit amplification
RpoB_fw	ACGTAAGCTTATCCAGGTCATCAAGGTCG	78
RpoB_rev	ACGTTCTAGAAGATCTTCTCGCAGAAGAG	77
RpoC_fw	ACGTAAGCTTGTGCTCGACGTCAACTTCT	73.3
RpoC3_rev	ACGTTCTAGATTACTGGTTGTACGGACCG	70.6
Primers for site-directed mutagenesis
2433_T458N_fw	AGTTCATGGACCAGAACAACCCGCTGTCGGG	80.31
2433_T458N_rev	CCCGACAGCGGGTTGTTCTGGTCCATGAACT	80.31
2433_A636S_fw	TTCCGCTGATCAAGTCGGAGTCGCCGCTGGTC	82.91
2433_A636S_rev	GACCAGCGGCGACTCCGACTTGATCAGCGGAA	82.91
2433_D486E_fw	GCGGGCCGGCCTGGAGGTCCGTGACGTGCAC	88.25
2433_D486E_rev	GTGCACGTCACGGACCTCCAGGCCGGCCCGC	88.25
2433_L555V_fw	AGGAGGACCGCTTCGTGATCGCGCAGGCCAAC	84.19
2433_L555V_rev	GTTGGCCTGCGCGATCACGAAGCGGTCCTCCT	84.19
2433_F593Y_fw	GCCGACGAGGTGGACTACATGGACGTCTCGCCG	85.35
2433_F593Y_rev	CGGCGAGACGTCCATGTAGTCCACCTCGTCGGC	85.35
2433_S624A_fw	CGCGCGCTCATGGGAGCGAACATGATGCGCC	84.28
2433_S624A_rev	GGCGCATCATGTTCGCTCCCATGAGCGCGCG	84.28
2433_L485F_fw	TGAGCGGGCCGGCTTCGACGTCCGTGACGTG	87.60
2433_L485F_rev	CACGTCACGGACGTCGAAGCCGGCCCGCTCA	87.60
2433_LD485FE_fw	TGAGCGGGCCGGCTTCGAGGTCCGTGACGTGC	83.70
2433_LD485FE_rev	GCACGTCACGGACCTCGAAGCCGGCCCGCTCA	83.70
2494_I467L_fw	CCCGCTGCTGGGCCTCACCCACAAGCGGCGTC	83.7
2494_I467L_rev	GACGCCGCTTGTGGGTGAGGCCCAGCAGCGGG	83.7
2494_N474S_fw	CACAAGCGGCGTCTGAGCTCGCTCGGCCCGGGT	91.3
2494_N474S_rev	ACCCGGGCCGAGCGAGCTCAGACGCCGCTTGTG	91.3
2494_S475A_fw	AGCGGCGTCTGAACGCGCTCGGCCCGGGTGG	85.0
2494_S475A_rev	CCACCCGGGCCGAGCGCGTTCAGACGCCGCT	85.0
2494_T638K_fw	CCGTGCCGCTGATCAAGGCCGAGGCCCCCCTCG	92.5
2494_T638K_rev	CGAGGGGGGCCTCGGCCTTGATCAGCGGCACGG	92.5
2494_A639S_fw	TGCCGCTGATCACCTCCGAGGCCCCCCTCGTC	81.8
2494_A639S_rev	GACGAGGGGGGCCTCGGAGGTGATCAGCGGCA	81.8
2494_NS474SA_fw	CAAGCGGCGTCTGACGGCGCTCGGCCCGGGTG	86.2
2494_NS474SA_rev	CACCCGGGCCGAGCGCCGTCAGACGCCGCTTG	86.2
SA_L463I_fw	CCCGCTCTCGGGAATCACCCACAAGCGCCGTC	80.6
SA_L463I_rev	GACGGCGCTTGTGGGTGATTCCCGAGAGCGGG	80.6
SA_S470N_fw	CACAAGCGCCGTCTGAACG CGCTCGGCCCGG	91.8
SA_S470N_rev	CCGGGCCGAGCGCGTTCAGACGGCGCTTGTG	91.8
SA_A471S_fw	AGCGCCGTCTGTCGTCGCTCGGCCCGGGTG	91.3
SA_A471S_rev	CACCCGGGCCGAGCGACGACAGACGGCGCT	91.3
SA_F484L_fw	CGTGAGCGGGCCGGCCTCGAGGTCCGTGACGT	88.03
SA_F484L_rev	ACGTCACGGACCTCGAGGCCGGCCCGCTCACG	88.03
SA_E485D_fw	AGCGGGCCGGCTTCGACGTCCGTGACGTGCAC	86.75
SA_E485D_rev	GTGCACGTCACGGACGTCGAAGCCGGCCCGCT	86.75
SA_FE485LD_fw	TGAGCGGGCCGGCCTCGACGTCCGTGACGTGC	78.66
SA_FE485LD_rev	GCACGTCACGGACGTCGAGGCCGGCCCGCTCA	78.66
SA_SA470NS_fw	CAAGCGCCGTCTGAACTCGCTCGGCCCGGGTG	82.4
SA_SA470NS_rev	CACCCGGGCCGAGCGAGTTCAGACGGCGCTTG	82.4
Primers for chimeric RpoB construction
NheSA_fw	ATCCGGCCGGTCGTCGCTAGCATCAAGGAGTTCTTC	78.33
NheSA_rev	GAAGAACTCCTTGATGCTAGCGACGACCGGCCGGAT	78.33
BspTISA_fw	ACGCCGGCGACGTGCTTAAGGCGGAGAAGGACGGTG	87.31
BspTISA_rev	ACCGTCCTTCTCCGCCTTAAGCACGTCGCCGGCGTC	87.31
Nhe_fw	CAGGTCGCTAGCATCAAGGAGTTCTTCG	61
BspTI_rev	CTCGGCCTTAAGGACGTCACCGGCGTCG	70

^aIntroduced mutations are underlined.