dna sequence of the bipa (bp1112) gene  (GenScript corporation)

Journal: Emerging Microbes & Infections

Article Title: The vaccine potential of Bordetella pertussis biofilm-derived membrane proteins

doi: 10.1038/emi.2014.58

Figure Lengend Snippet: Biofilm-associated proteins and vaccine candidate selection criteria

Article Snippet: The entire DNA sequence of the bipA (BP1112) gene lacking the N-terminal signal sequence (amino acids 1–20) and membrane spanning domain (amino acids 21–66) was codon optimized for expression in E. coli , synthesized by GenScript (GenScript USA Inc. Piscataway, NJ, USA), and cloned into pET28-TEVsite vector (modified from pET28a vector; Novagen, Madison, WI, USA) to generate an N-terminally His 6 -tagged BipA protein (rBipA). rBipA expression from the pET28-TEVsite vector was induced in E. coli OverExpress C41(DE3) (Lucigen) cultured to OD 600 =0.6–0.7 at 37°C by addition of 1 mM isopropyl β- d -1-thiogalactopyranoside for 3 h. Induced cultures were harvested by centrifugation (4000 g for 15 min at 4°C) and enzymatically lysed by resuspending the pellet in Bacterial Protein Extraction Reagent (Thermo Scientific, Waltham, MA, USA) containing 25 U/mL benzonase nuclease (Novagen), 100 μg/mL lysozyme, and protease inhibitors (Roche, Mannheim, Germany), and incubating for 30 min at room temperature.

Techniques: Selection, Quantitative Proteomics, Membrane