Table 2.

Primers used in this study for construction of deletion mutants and real-time PCR

Italic type indicates the restriction site integrated into the primer.

Primer purpose	5′-end amplicon		Deleted gene/amplicon/probe*	3′-end amplicon
	Primer	Nucleotide sequence (5′→3′)		Primer	Nucleotide sequence (5′→3′)
General
	lrgA-A	GCAATTGGCACATCCTCCAC	lrgA	lrgA-SphI-C	CTATCAAAAGCATGCATGTGGCAAG
	lrgA-SphI-B	CTGAAATAAGCATGCAAACGAGCAG		lrgA-D	CCATGGCAGTGATGGCAGTA
	lrgB-A	GCAATCGGGGACAGTTTTGA	lrgB	lrgB-EcoRI-C	GCAGCCTTTGAATTCGAATTAGGAG
	lrgB-SalI-B	GCAAAGAAAGTCGACTGTAAGAGAA		lrgB-D	ACAGACCGCTTTGAGGTTGC
	lytS-A	ACTGAACAGCCAGTGCACCA	lytS	lytS-BamHI-C	GCAGTGCTAGGATCCTACACTTTGA
	lytS-BamHI	GCCAGAATCGGATCCATACCAAGTC		lytS-D	TCAAAACTGTCCCCGATTGC
	cidA-A	TGCGGTCAGTTTTGCTGTGG	cidA	cidA-BamHI-C	GAGACATTAGGATCCAGACTTTCCA
	cidA-BamHI-B	CAAATTCGCGGATCCAAGAAAAGAG		cidA-D	TGAGACAAAAGTGTTCCCAACC
	cidB-A	GCGCTTTTCAGGCAAGCAGA	cidB	cidB-BamHI-C	ACGGGTTTGGGATCCGTCTTTGTAT
	cidB-BamHI-B	TAGGCAAATGGATCCAGCCAAAGAC		cidB-D	TGGCGCCAAATCTCTTACGC
RT-PCR†or real-time RT-PCR
	lrgA-sense	TTGCCTAAAGCCTTACCGATTCC	lrgA	lrgA-antisense	GCCTGATGGGACAAACATAAAGC
	lrgB-sense	GGCAAAAGGATTGGGAACTGATG	lrgB	lrgB-antisense	TGGAACGGCAAAGGCAATGG
	lytS-sense	TTGTCAGTTCTGCTTTGGTAGG	lytS	lytS-antisense	CAATGACCTGCGAAGTAGATGG
	lytT-sense	CATCCTCCACTTGTCGTCTTTGC	lytT	lytS-antisense	CACACGCCCCTGCTCAAAAG
	cidA-sense	ATCCGTTTGCGTCATATCAATGC	cidA	cidA-antisense	CCATAATCCCCACTGCTGCTG
DIG-probe synthesis
	lrgA-sense	CACAATCAAAATCAGCACCT	lrgA	lrgA-antisense	TCACCTTTTTGATAGACAGAA
	cidB-sense	TTTTTCGAATCCTCTTTTTG	cidB	cidB-antisense	CAACAACAACCAGTGTTACG

*Additional primers used for deletion of lrgAB were as follows: lrgA-BamHI-B, CTGAAATAAGGATCCAAACGAGCAG; lrgB-BamHI-C, GCAGCCTTTGGATCCGAATTAGGAG.

†Primer sets for RT-PCR were employed as follows: lrgA-sense/lrgB-antisense primers for lrgA–lrgB, and lrgB-sense/lytS-antisense primers for lrgB–lytS.