Journal: Applied and Environmental Microbiology

Article Title: A Counterselectable Sucrose Sensitivity Marker Permits Efficient and Flexible Mutagenesis in Streptococcus agalactiae

doi: 10.1128/AEM.03009-18

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: Strains E. coli DH5α Chemically competent cloning strain New England Biolabs item no. C2987H B. subtilis 168 Legacy strain derived from B. subtilis Marburg 39 GBS CNCTC 10/84 Serotype V, sequence type 26 40 GBS A909 Serotype Ia, sequence type 7 41 GBS AR1598 Serotype V, sequence type 26 10/84 mutant with barcoded premature stop codon in cylE gene This study Plasmids pDC123 Shuttle vector with the chloramphenicol acetyltransferase gene used as PCR template 42 pHY304 Broad-host-range, temperature-sensitive mutagenesis plasmid; pWV01 derivative; erythromycin resistant 12 pMBsacB pHY304 with p23- sacB cassette cloned into AvaII site, for sucrose-counterselectable GBS mutagenesis This study pORI23 Shuttle vector with ori ColE1 and Lactococcus -derived Gram-positive origin of replication; erythromycin resistant; p23 promoter adjacent to BamHI restriction site 43 pSacB23 pORI23 with sacB cloned into BamHI site for expression off p23 promoter This study Open in a separate window Strains and plasmids used in this study Cloning technique.

Techniques: Plasmid Preparation, Cloning, Derivative Assay, Sequencing, Mutagenesis, Clone Assay, Expressing