Cloning, Expression and Purification of Chalcone Synthase from Solanum Tuberosum
نویسندگان
چکیده
Chalcone synthase (CHS) gene was amplified by PCR from cDNA of Solanum tuberosum leaf tissue. The gene was cloned into pGEMT easy vector and sequenced. It was then ligated into pET 30 vector to construct recombinant plasmid pET30-CHS. The recombinant was transformed into E.coli BL21 strain and the target gene was over expressed using IPTG. Optimum condition of IPTG concentration and temperature for over expression was found to be 1 mmol L -1 and 37oC respectively. The results showed that the His-tagCHS was highly heterogonous expressed in the inclusion bodies into the host cell. However, the His-tagCHS fusion protein was purified using affinity chromatography with Ni 2+ -NTA resin column. SDS-PAGE analysis demonstrated that the host with pET30-CHS generated a 42 kD His-tag-CHS fusion protein.
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