Title: Prokaryotic Expression and Purification of GP73
Abstract: Objective To prokaryotically express and purify Golgi protein 73(GP73). Methods We amplified GP73 gene from human hepatoma cell HepG2 by RT-PCR and cloned it into the prokaryotic expression vector pET21a(+)-TRX, then transformed it into Escherichia coli BL21(DE3). The expression of GP73 protein was induced by isopropyl β-D-1-thiogalactopyranoside(IPTG) and purified by His-tag magnetic beads. SDS-PAGE was used to identify GP73. Results Gene GP73 was cloned from HepG2 without any mutation. The recombinant plasmid pET21a(+)-TRX-GP73 was constructed correctly and the recombinant protein GP73 was expressed successfully with an expected molecular weight of 80KD. Conclusion We successfully expressed and purified the recombinant protein GP73 in Escherichia coli BL21(DE3) and layed a foundation for following study.
Publication Year: 2014
Publication Date: 2014-01-01
Language: en
Type: article
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