Journal: Life

Article Title: Characterization and PCR Application of Family B DNA Polymerases from Thermococcus stetteri

doi: 10.3390/life14121544

Figure Lengend Snippet: Expression and purification assay of Tst DNA polymerases. ( A ) Expression analysis of Tst(wt) and the TstP36H–Sso7d DNA polymerase gene in E. coli Rosetta 2(DE3) pLysS cells containing the recombinant plasmids. A. Lane M, protein molecular mass markers; Lane N, sonicated pellet (N P ) and lysate (N L ) of uninduced cells; Lane I, sonicated pellet (I P ) and lysate (I L ) of induced cells. ( B ) Purification of Tst(wt) and TstP36H–Sso7d DNA polymerases. Lines 1–3 indicate protein-containing fractions from chromatographies in a HiTrap Heparin™ column, Line 4, resulting in the Heparin-purified Tst(wt) DNA polymerase, Line 5, resulting in the phenyl-purified TstP36H–Sso7d DNA polymerase.

Article Snippet: E. coli strain DH5a (Thermo Scientific, Waltham, MA, USA) was used for plasmid propagation while E. coli strain Rosetta2 (DE3)pLysS (Novagen, Merck KGaA, Darmstadt, Germany) was used for gene expression.

Techniques: Expressing, Purification, Recombinant, Sonication