Journal:
Article Title: The Alternative Sigma Factor ? B of Bacillus cereus : Response to Stress and Role in Heat Adaptation
doi: 10.1128/JB.186.2.316-325.2004
Figure Lengend Snippet: Overproduction and purification of σB. (Left panel) SDS-PAGE of protein extracts from E. coli BL21-Codonplus-(DE3)-RIL carrying either pET28-b or pMT01. (Right panel) Immunodetection of σB with anti-σB antiserum. Ten micrograms of protein was loaded for each sample. Lane 1, crude protein extract from E. coli carrying pET28-b; lane 2, crude protein extract from E. coli carrying pMT01; lane 3, crude protein extract from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 4, inclusion bodies isolated from E. coli carrying pMT01 after induction with 1 mM IPTG for 2 h; lane 5, representative fraction of purified σB after elution from an Ni2+ affinity column. The arrow indicates the position of the overproduced σB protein.
Article Snippet: The plasmids used in this study are listed in Table . table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Plasmid Relevant properties Source or reference pGEM-T PCR cloning vector, Amp r Promega pET28-b E. coli overexpression vector, Kan r Novagen pMT01 pET28-b derivative containing sigB under control of the T7 RNA polymerase promoter This study pUC18ERY Amp r Ery r 45 pATΔS28 tra + conjugative suicide vector for B. cereus group, Spc r 32 pATΔS28ery pATΔS28 derivative containing erythromycin resistance cassette from pUC18ERY, Spc r Ery r This study pATΔS28eryBY pATΔS28ery derivative containing 1.1-kb downstream flanking region of sigB , Spc r Ery r This study pATΔsigB pATΔS28ery derivative containing 1.1-kb upstream and downstream flanking regions of sigB , Spc r Ery r This study Open in a separate window Plasmids used in this study
Techniques: Purification, SDS Page, Immunodetection, Isolation, Affinity Column
Addgene inc is a verified supplier 
