Journal: Applied and Environmental Microbiology

Article Title: Involvement of Two-Component System CBO0366/CBO0365 in the Cold Shock Response and Growth of Group I (Proteolytic) Clostridium botulinum ATCC 3502 at Low Temperatures

doi: 10.1128/AEM.00555-12

Figure Lengend Snippet: Oligonucleotide primers

Article Snippet: Bacterial strains and plasmids table ft1 table-wrap mode="anchored" t5 caption a7 Primer Sequence (5′→3′) a Use c Binding site in ATCC 3502 genome b cbo0365 -f AATGATGCCTAAGATGGATGGT qRT-PCR 424216–424237 cbo0365 -r TCTGCACCTGTGGTTAATCCT qRT-PCR 424324–424344c cbo0366 -f GGCATACCAAGAGCAGAAACCA qRT-PCR 425289–425310 cbo0366 -r ATTTGCAGCAAGCCCTTTGA qRT-PCR 425421–425440c 16S rrn -f TTGTCGTCAGCTCGTGTCGT qRT-PCR 10290–10309 16S rrn -r CCTGGACATAAGGGGCATGA qRT-PCR 10431–10450c cbo0366 -f-NdeI NNNNNN CATATG AAACCTTTCAAATATATA Construction of overexpression plasmid for cbo0366 424779–424796 cbo0366- r-NheI NNNNNN GCTAGC TTATTCATCCTCTGCCATAA Construction of overexpression plasmid for cbo0366 426235–426254c EBS Universal CGAAATTAGAAACTTGCGTTCAGTAAAC Retargeting of pMTL007, control for correct mutation site in cbo0365 and cbo0366 NA cbo0365 -IBS AAAAAAGCTTATAATTATCCTTAAAAGACATCAGAGTGCGCCCAGATAGGGTG Construction of pMTL007- cbo0365- 48s NA cbo0365 -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCATCAGAGATAACTTACCTTTCTTTGT Construction of pMTL007- cbo0365- 48s NA cbo0365 -EBS2 TGAACGCAAGTTTCTAATTTCGGTTTCTTTCCGATAGAGGAAAGTGTCT Construction of pMTL007- cbo0365- 48s NA cbo0365 M-f TTGTTGATGATGAAAAAGAAATCA Control of pMTL007- cbo0365- 48s 424074–424097 cbo0366 -IBS AAAAAAGCTTATAATTATCCTTAACTATCCAAGAAGTGCGCCCAGATAGGGTG Construction of pMTL007- cbo0366- 267s NA cbo0366 -EBS1d CAGATTGTACAAATGTGGTGATAACAGATAAGTCCAAGAACTTAACTTACCTTTCTTTGT Construction of pMTL007- cbo0366- 267s NA cbo0366 -EBS2 TGAACGCAAGTTTCTAATTTCGATTATAGTTCGATAGAGGAAAGTGTCT Construction of pMTL007- cbo0366- 267s NA cbo0366 M-f TTTTTAATGGCTATTATGACCTTTATT Control of pMTL007- cbo0366- 267s 424866–424892 Open in a separate window a Underlined portions of sequences indicate the restriction endonuclease site. b Based on EMBL accession number {"type":"entrez-nucleotide","attrs":{"text":"AM412317","term_id":"148287495","term_text":"AM412317"}} AM412317 (positions are shown according to base pair number). c, complement strand; NA, not applicable. c 48s and 267s represent mutations between bases 48 and 49 and between bases 267 and 268, respectively, in the sense orientation.

Techniques: Sequencing, Binding Assay, Over Expression, Plasmid Preparation, Mutagenesis