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Table 2 Primers used in this study

From: Investigating the role of small, acid-soluble spore proteins (SASPs) in the resistance of Clostridium perfringens spores to heat

Primers name

Primer Sequencea

Positionb

Gene

Usec

CPP 7

5' GCTTACAAATTACCAAAGCC 3'

-36 to -17

ssp1

PCR, Se

CPP 8

5' CAGTATTAGCGAAAGGTTTG 3'

83 to 202

ssp1

PCR, Se

CPP 9

5' CTCCTATAATTCCCTCTCAT 3'

-105 to -86

ssp2

PCR, Se

CPP10

5' GTAGACTTTAATAGGTTCAGG 3'

180 to 200

ssp2

PCR, Se

CPP11

5' CTGCACATCATAATATTGAAAGG 3'

-80 to -58

ssp3

PCR, Se

CPP 13

5' GAGGGTCCTATTGTAGGAGGATT 3'

-2010 to -1888

ssp3

MP, SB

CPP 16

5' ATAGCAGGAGGAGCTATTCCAC 3'

2099 to 2120

ssp3

MP, SB

CPP 21

5' CGGAGACCTTAGCTCAAGTTAACGCGGAAGCGTTGGTGGAG 3'

95 to 135

ssp3

SDM

CPP 22

5' CTCCACCAACGCTTCCGCGTTAACTTGAGCTAAGGTCTCCG 3'

95 to 135

ssp3

SDM

CPP 34

5' GCTATGGATCTTATGGAAGG 3'

423 to 442

ssp3

PCR, Se

CPP37

5' CGGCTTCTAGCACATCTTCT 3'

-1662 to -1593

ssp2

MP, SB

CPP38

5' TATGTGGAGCAGGAATTGCC 3'

1449 to 1469

ssp2

MP, SB

CPP47

5'GTACCATTCTCAGATTACAATGTCGCGAAGCGTTGGTGGAGAAATGG3'

73 to 113

ssp2

SDM

CPP48

5' CCATTTCTCCACCAACGCTTCGCGACATTGTAATCTGAGAATGGTAC 3'

73 to 113

ssp2

SDM

CPP45

5' CCAGGAAAGTATGGACAAGC

-1564 to -1544

ssp1

MP

CPP139

5' CCTCACCATTATCCTCTACAAG 3'

1782 to 1802

ssp1

MP

CPP104

5' GCTAGAGAATTAGGGGTACCTCGCGACCTAAGTTCAAGACAA 3'

60 to 116

ssp1

SDM

CPP105

5' TTGTCTTGAACTTAGGTCGCGAGGTACCCCTAATTCTCTAGC 3'

60 to 116

ssp1

SDM

CPP 57

5' GCGTCGAC CTGTTTGAGCTTTTTTC 3'

-200 to -213

ssp3 promoter

GUS

CPP 58

5' GCTGCAG CCTGGAACTAAATGTTGT 3'

3 to 21

ssp3 promoter

GUS

CPP 63

5' GCGTCGAC TAGGTGCAGAAGC 3'

-219 to -203

ssp1 promoter

GUS

CPP 64

5' GCTGCAG CTGGAACTAATGATTTTGAC 3'

3 to 21

ssp1 promoter

GUS

CPP 65

5' GCGTCGAC GGAACTAAAGCTAAATTTGG 3'

-238 to -223

ssp2 promoter

GUS

CPP 66

5' GCTGCAG GCTTCTGGTACTAAATGTTGTG 3'

3 to 25

ssp2 promoter

GUS

  1. a Restriction sites are marked by underlining.
  2. bThe nucleotide position numbering begins from the first codon and refers to the relevant position within the respective ssp gene sequence [18].
  3. cPCR, polymerase chain reaction; Se, sequencing studies; MP, construction of mutator plasmid; SB, southern blot analysis; SDM, site-directed muatgenesis. GUS, construction of plasmid for β-glucuronidase assay.