ClpP participates in stress tolerance, biofilm formation, antimicrobial tolerance, and virulence of Enterococcus faecalis

Table 4 Primers used for △clpP mutant construction and complemented strains

Primers^a	Sequences (5′ → 3′)	Location^b	PCR product size (bp)	Note^c
Construction of ΔclpP mutant
clpP us-F	GCTCTAGATTTGGGGTGTTTGTTTAGCAG	530,796–530,816	1059	XbaI
clpP us-R	CGGGATCCGGAAGTAAATCCTCCTATATAAAG	529,774–529,794		BamHI
clpP ds-F	CGGGATCCCAGCGACACTCGCTGTTTCCA	529,139–529,159	1044	BamHI
clpP ds-R	AACTGCAGAGGCGTAGATGAACCAGTGGT	528,132–528,152		PstI
Construction of ΔclpP complemented strain
HBclpP-F	CGGGATCCAGGCATTCAAAGTGCTTTGTG	529,816–529,836	714	BamHI
HBclpP-R	CCCTCGAGTGGAAACAGCGAGTGTCGCTG	529,139–529,159		XhoI
RT-qPCR
recA-F	CGACTAATGTCTCAAGCACTAC	2,574,587–2,574,608	106
recA-R	CGAACATCACGCCAACTT	2,574,503–2,574,520
clpP-F	TTAATTCCAACAGTTATTGAA	529,746–529,766	198
clpP-R	ACCAGGAGAGTTAATGTA	529,569–529,586

^aPrimers were designed according to the genomic sequence of E. faecalis OG1RF (GenBank accession number CP002621.1). F, forward primer; R, reverse primer. ^bLocation of the primer in the genomic sequence of E. faecalis OG1RF (GenBank accession number CP002621.1). ^cThe underlined sequences represent restriction enzyme sites

ISSN: 1471-2180