Cloning and expression of egI gene from Penicillium decumbens L-06
斜卧青霉L-06内切葡聚糖酶Ⅰ基因的克隆与表达

Objective] The research focus on cloning endoglucanase I (egI) gene from Penicillium decumbens L-06 and expressing in Escherichia coli with high efficiency. Methods] egI gene was cloned from Penicillium decumbens L-06 by RT-PCR method. Recombinant plasmid pET32a-egI was constructed and was transformed into Escherichia coli rosetta(DE3). Recombinant protein with His-tag was expressed in E. coli rosetta(DE3) after induction with IPTG and then was purified with the Ni-NTA affinity chromatography. Results] As expected, the relative molecular mass was approximately 80kD after analyzed by SDS-PAGE and Western blotting. Hydrolysis activity of recombinant protein was assayed by cellulase activity staining and DNS method (2.56 IU/mL). Conclusion] The results achieve the purpose as constructing prokaryotic expression system and expressing egI gene.