LsrB-based and temperature-dependent identification of bacterial AI-2 receptor

AMB Express

Table 1 Primers used in this study

Primers	Oligonucleotide sequence (5′–3′)	Description	Product size (bp)
LuxS-UF	TTCTGATGCGCTGTTACGT	The upstream sequence of luxS	845
LuxS-Overlap-UR	TCGGCAGTGCCGCGTCGACGGGGTGTTCATTGTTTTCG^a
LuxS-Overlap-DF	TGAACACCCCGTCGACGCGGCACTGCCGAAAGAGAAG	The downstream sequence of luxS	884
LuxS-DR	TGACCGACGATAACCCGA
pkD4-Kan-F	CGCGTCGACTGTAGGCTGGAGCTGCTT	Kanamycin resistance cassette	1494
pkD4-Kan-R	CGCGTCGACCATATGAATATCCTCCTTAGTTC
LsrB-F	CGCGGATCCATGGCAAGACACAGCATTAAAAT^b	lsrB	1023
LsrB-R	CCCAAGCTTTCAGAAATCATATTTGTCGATATTG^c
LuxS-F	CGCGGATCCATGCCGTTGTTAGATAGCTT^b	luxS	516
LuxS-R	CCCAAGCTTCTAGATGTGCAGTTCCTGCA^c
LuxS-OutF	GCGATTTGTTCTTCTTTCCTG^d	Primers for identification of luxS deletion	2519
LuxS-OutR	GATCAAGAATCGTCACAGG
LuxS-InF	GAAGTGATGCCAGAAAGAGGG^e	Primers for identification of luxS deletion	307
LuxS-InR	CCAGAATGCTACGCGCAATAT

^{a, b, c} SalI, BamHI and HindIII restriction sites are underlined, respectively
^dA 307 bp PCR product was amplified from wild-type strain BL21(DE3), and no product was amplified from mutant strain BL21ΔluxS, using primers LuxS-inF/LuxS-inR
^eA 2519 or 2209 bp PCR product was amplified from wild-type strain BL21(DE3), or mutant strain BL21ΔluxS without kan, respectively, using primers LuxS-OutF/LuxS-OutR