Lsr operon is associated with AI-2 transfer and pathogenicity in avian pathogenic Escherichia coli

Zuo, Jiakun; Yin, Huifang; Hu, Jiangang; Miao, Jinfeng; Chen, Zhaoguo; Qi, Kezong; Wang, Zhihao; Gong, Jiansen; Phouthapane, Vanhnaseng; Jiang, Wei; Mi, Rongsheng; huang, Yan; Wang, Chen; Han, Xiangan

doi:10.1186/s13567-019-0725-0

Veterinary Research

Table 2 Primers used in this study

From: Lsr operon is associated with AI-2 transfer and pathogenicity in avian pathogenic Escherichia coli

Primers	Oligonucleotide sequence (5′ to 3′)	Description	Product size (bp)
lsrK-F	CTATAACCCAGGCGCTTTCCATA	Partial sequence of lsrK	1593
lsrK-R	ATGGCTCGACTCTTTACCCTTTC	Partial sequence of lsrK	1593
lsrR-F	TTAACTACGTAAAATCGCCGCTG	Partial sequence of lsrR	954
lsrR-R	ATGACAATCAACGATTCGGCAAT	Partial sequence of lsrR	954
LsrA-F	ATGCAAACGAGTGATACCCGCGC	Partial sequence of lsrA	1539
lsrA-R	ACTTCAGCATGACGCCTCCTGAC	Partial sequence of lsrA	1539
lsrC-F	CTGAAGTTTATTCAGAACAACCGTG	Partial sequence of lsrC	826
lsrC-R	TATCGATCTGCGTCAGGAACCAT	Partial sequence of lsrC	826
lsrD-F	GCCGCATAATGCGTATTCGCTAC	Partial sequence of lsrD	1003
lsrD-R	CTTTATGGCAATGGGTTATTGGC	Partial sequence of lsrD	1003
lsrB-F	ATGACACTTCATCGCTTTAAGA^a	Partial sequence of lsrB	826
lsrB-R	CTGAAATTTTGCCTTGCTGAAC^a	Partial sequence of lsrB	826
lsrF-F	GTAAAGATTTTCGTACCGATCAAC	Partial sequence of lsrF	657
lsrF-R	TTTTACCGCCAGCAATAACAATG	Partial sequence of lsrF	657
lsrG-F	ATGCACGTCACACTGGTTGAAAT	Partial sequence of lsrG	291
lsrG-R	TCACGGCATCAAACCATTGAACA	Partial sequence of lsrG	291
APEC94lsr-F	TCCCTTGAATATCGTACTGG	Using for DNA sequencing of lsr operon
APEC94lsr-R	TGCGCTTCGATGTCTTAC	Using for DNA sequencing of lsr operon
pCold-F1	CCCAAGCTTGAAATCGTATTTGCCGAT	Using for identification of lsrB-APEC recombinant plasmid	1150
pCold-R	GGCAGGGATCTTAGATTCTG	Using for identification of lsrB-APEC recombinant plasmid	1150
lsr-UF	TCTAAAAGAAGGGAAATAAG^b	The upstream sequence of lsr operon	763
lsr-UR	CCAGCCTACA GCATGGTACAAGCTGAATC	The upstream sequence of lsr operon	763
lsr-CF	TGTACCATGCTGTAGGCTGGAGCTGCTT	Chloramphenicol resistance cassette (cat)	1013
lsr-CR	CATTGAACAGCATATGAATATCCTCCTTAGTTC	Chloramphenicol resistance cassette (cat)	1013
lsr-DF	TATTCATATGCTGTTCAATGGGTTGATGC	The downstream sequence of lsr operon	638
lsr-DR	TGGTGATAGTAGGTGGTTCG^b	The downstream sequence of lsr operon	638
PstI-lsr-CF	AAAACTGCAGGGTCTGTATTGAGTGTTAGTTGGAGGTGGG^c	The lsr operon complement sequence	9440
PstI-lsr-CR	AAAACTGCAGCAGTTGCGGATGTCTGCTT^c	The lsr operon complement sequence	9440
LsrB-BamHI-F	CGCGGATCCATGACACTTCATCGCTTTA^d	The ORF sequence of lsrB	1023
LsrB-HindIII-R	CCACTTTCAACGAGCTGATG^e	The ORF sequence of lsrB	1023
fimH-RTF	GTGCCAATTCCTCTTACCGTT	Partial DNA sequence of fimbrial protein FimH	165
fimH-RTR	TGGAATAATCGTACCGTTGCG	Partial DNA sequence of fimbrial protein FimH	165
fliD-RTF	TTCAGACGCAGTTGAAATCG	Partial DNA sequence of flagellar filament capping protein FliD	184
fliD-RTR	TTACCATCGCCAACAATCAA		184
flhA-RTF	CGGCATCGTACTCTGGAACT	Partial DNA sequence of flagellar biosynthesis gene flhA	172
flhA-RTR	CATTACCCATCCGTTCGTTC	Partial DNA sequence of flagellar biosynthesis gene flhA	172
dosP-RTF	ATAAACCCACGCCCATATCA	Partial DNA sequence of oxygen-sensing cyclic-di-GMP phosphodiesterase DosP	194
dosP-RTR	GGCGTTATCCGTGAACTTGT		194
pgaA-RTF	GGCAATGGTCTCCTTGTGAT	Partial DNA sequence of poly-beta-1,6 N-acetyl-d-glucosamine export porin PgaA	195
pgaA-RTR	GATCATCTTGGCGCGTTATT		195
fhuD-RTF	AACTATCGCCTGTGGGTCAG	Partial DNA sequence of iron-hydroxamate transporter substrate-binding subunit fhuD	153
fhuD-RTR	TGCCAGCATTTCTGATGAAG		153
dnaE-RTF	GATTGAGCGTTATGTCGGAGGC	Partial DNA sequence of the internal control dnaE	80
dnaE-RTR	GCCCCGCAGCCGTGAT	Partial DNA sequence of the internal control dnaE	80

^aA 826-bp PCR product was amplified from wild-type strain APEC94 but no PCR product was obtained from mutant strain APEC94Δlsr(Cm) using primers lsrB-F/lsrB-R.
^bA 2414-bp PCR product was amplified from mutant strain APEC94Δlsr(Cm) but no product was amplified from wild-type strain APEC94 in a limited amplification time using primers lsr-UF/lsr-DR.
^cPstI restriction sites are underlined.
^dBamHI restriction sites are underlined.
^eHindIII restriction sites are underlined.

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ISSN: 1297-9716

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