Journal:

Article Title: The Flagellar Muramidase from the Photosynthetic Bacterium Rhodobacter sphaeroides ▿

doi: 10.1128/JB.01073-07

Figure Lengend Snippet: Bacterial strains, plasmids, and oligonucleotides

Article Snippet: The bacterial strains, plasmids, and oligonucleotides used in the present study are described in Table . table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain, plasmid, or oligonucleotide Relevant characteristics or sequence a Source or reference Strains E. coli JM103 hsdR4 Δ( lac - pro ) F′ traD 36 proAB lacI q Z ΔΜ15 3 M15[pREP4] Lac − Ara − Gal − Mtl − F′ RecA + Uvr + Lon + QIAGEN BL21DE3pLysS F′ ompT hsdSB ( r B − m B − ) gal dcm (DE3)pLysS Novagen S17-1 recA endA thi hsdR RP4-2-Tc::Mu::Tn7; Tp r Sm r 45 R. sphaeroides WS8-N Wild type; spontaneous Nal r 47 SltF1 WS8 sltF Δ 1 :: aadA derivative; Fla − Spc r This study Plasmids pQE30 Expression vector; Ap r N-terminal His 6 tag QIAGEN pQE60 Expression vector; Ap r C-terminal His 6 tag QIAGEN pRSJ flgJ cloned into the NcoI/BglII sites of pQE60 sites This study pRS0072 RSP0072 cloned into NcoI/BamHI sites of pQE30 sites This study pRS0072E57A RSP0072 E57A cloned into pQE30 This study pRS0072E83A RSP0072 E83A cloned into pQE30 This study pJQ200mp18 Sucide vector used for gene replacement 42 pTZ19R Cloning vector, pUC derivative, Ap r Pharmacia pRK415 pRK404 derivative; used for expression in R. sphaeroides 24 pWM5 pUC derivative carrying the omega-Spc r cassette 34 pBG313 3.8-kb BamHI fragment carrying RSP0072 from WS8-N Cloned into pTZ19R 16 pRKsltF 1.4-kb PstI/PstI fragment from pBG313 subcloned into pRK415 carrying sltF (RSP0072); Tc r This study Oligonucleotides F1 5′-CTGATCTAGACCCTCCGGCCCCGGCCACGGTG-3′ This study R2 5′-CCGGAATTCGACGAGCGCGAACCTCATGCC-3′ This study F3 5′-CGGGAATTCCTCAACTGGCGCTGGCATGCG-3′ This study R4 5′-CTGATCTAGAGGGGTCACGCCCGAGAGACAG-3′ This study SPC1 5′-CCTGGATATCGGGCAGATCCGTGC-3′ This study SPC2 5′-TCATGATATCTCTCCCAATTTGTG-3′ This study orf72F 5′-CATGGAGCTCGCGGACGAGGGCTGCGAGACG-3′ This study orf72R 5′-CCCGAAGCTTTCACGGTTGCATTGCGAGCAG-3′ This study orf72F57A 5′-GGCGATTGCCCGCGTGGCGTCGGGCCGGGGCGGGC-3′ This study orf72R57R 5′-GCCCGCCCCGGCCCGACGCCACGCGGGCAATCGCC-3′ This study orf72F83A 5′-CTTCGAGACCCGGGCCGCGGCGGTGCGCATGCTC-3′ This study orf72R83A 5′-GAGCATGCGCACCGCCGCGGCCCGGGTCTCGAAG-3′ This study JF 5′-CATGCCATGGATCTGAAGCTTCAGTCC-3′ This study JR 5′-GGAAGATCTCGACTTGCCGTCCCTGACGAGAG-3′ This study Open in a separate window a Spc r , spectinomycin resistance; Tc r , tetracycline resistance; Tp r , trimethoprim; Ap r , ampicillin resistance; Sm r , streptomycin resistance; Nal r , nalidixic acid resistance.

Techniques: Plasmid Preparation, Sequencing, Acetylene Reduction Assay, Expressing, Clone Assay, Cloning