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Table 2 Primers used in the present study

From: A previously uncharacterized gene stm0551 plays a repressive role in the regulation of type 1 fimbriae in Salmonella enterica serotype Typhimurium

Purpose and name

Sequence (5′ to 3′)

Description

Construction of the stm0551 mutant

stm0551pKD13-F

GCTCTGATGTTTCAATGCCTTCCATCAGC ATTAACTGATTCCGGGGATCCGTCGACC

Annealing Temp.: 55°C; amplicon length: 1,550 bp

stm0551pKD13-R

GGCACAGGGTATTTTGTTAAAGGAAAGG ATAAATCCCTGTAGGCTGGAGCTGCTTCG

 

Cloning

stm0551-F

GGATCC CATCCTGCTTTTTCCATTGCTCTAATAT

Bam HI restriction site (underlined)

stm0551-R

GATATC ACTCACTTAACTTTTTACAAGGCTTACG

Eco RV restriction site (underlined)

  

Annealing Temp.: 55°C; amplicon length: 500 bp

Construction of the fusion protein

stm0551-TOPO-F

CACCATGGTGGCACAGGGTATTTTGTTAA

Annealing Temp.: 50°C; amplicon length: 316 bp

stm0551-TOPO-R

ATATATATCTGGTAATATGGCTGG

 

fimY-TOPO-F

CACCATGCGCAGCGTACCACGCAG

Annealing Temp.: 50°C; amplicon length: 727 bp

fimY-TOPO-R

AAAAATGTCGTGGAAAGTAACGT

 

E49A-TOPO-F

ATCGGCTATGC GGTCCTGACGCAACTTCCG

Mutation site (underlined)

E49A-TOPO-R

CGGAAGTTGCGTCAGGACCG CATAGCCGAT

Mutation site (underlined)

RT-PCR analysis

fimA-RT-F

ACTATTGCGAGTCTGATGTTTG

 

fimA-RT-R

CGTATTTCATGATAAAGGTGGC

 

fimZ-RT-F

ATTCGTGTGATTTGGCGT

 

fimZ-RT-R

ACTTATCCTGTTGACCTT

 

fimY-RT-F

GAGTTACTGAACCAACAGCT

 

fimY-RT-R

GCCGGTAAACTACACGATGA

 

fimW-RT-F

AAAGTGAAAGTAAAGCGG

 

fimW-RT-R

AAGAGATAGATAATGCCCG

 

stm0551-RT-F

GCCATAAATAACCTTGTTCC

 

stm0551-RT-R

CATTCATATCTCAACAGCGA

 

16 s-F

TTCCTCCAGATCTCTACGCA

 

16 s-R

GTGGCTAATACCGCATAACG

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