Optimized production of a biologically active Clostridium perfringens glycosyl hydrolase phage endolysin PlyCP41 in plants using virus-based systemic expression

摘要

Protein production in . Bacterial cultures containing pET21a: Lysin D or pET21a: PlyCP41 (PlyCP41) were induced by addition of IPTG and proteins were purified using the BugBuster reagent. Total (T), soluble (So), and inclusion body (IB) fractions were collected. Five μl aliquots were run on a protein gel. Purification with PlyCP41 with Ni-NTA columns under native conditions. CP41, Soluble fraction from BugBuster fraction added to the Ni-NTA column; FT, flow through; W1, Wash 1; W2, Wash 2; E1, Elute 1; E2, Elute 2, E3, Elute 3; E4, Elute 4; E5, Elute 5; E6, Elute 6. Both gels were stained with SimplyBlue Safe Stain. M = Precision Plus Kaleidoscope protein standards