Journal: Microbial Cell Factories
Article Title: Revisiting Escherichia coli as microbial factory for enhanced production of human serum albumin
Figure Lengend Snippet: Cloning of HSA . a 1% agarose gel showing amplified PCR product and pET23b vector. Lane M, 1 kb DNA ladder; Lane 1, Uncut pET23b plasmid; Lane 2, a double digested pET23b vector with Nhe I and Xho I restriction enzymes; Lane 3, PCR amplified a product of HSA gene at an optimized annealing temperature of 58 °C. b 1% agarose gel showing the release of HSA gene after double digestion of constructed pETHSA vector with Nhe I and Xho I restriction enzymes. Lane M, 1 kb DNA ladder; Lane 1, Undigested pETHSA plasmid; Lane 2, digested pETHSA construct. c 12% SDS PAGE showing expression of rHSA in different strains of E. coli . Lane UI, Uninduced cells; Lane 1, Induced BL21 (DE3) E. coli cells; Lane 2, Induced Rosetta (DE3) E. coli cells; Lane 3, Induced Origami2 (DE3) E. coli cells
Article Snippet: The PCR fragments were double-digested with Nhe I and Xho I and then subcloned into a pET23b expression vector (Novagen, USA) that had been pre-digested with the same enzymes.
Techniques: Clone Assay, Agarose Gel Electrophoresis, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Construct, SDS Page, Expressing