1. Department of Endocrinology，Affiliated Hospital of Jiangsu University，Zhenjiang Jiangsu 212001;2.Institute of Life Sciences, Jiangsu University, Zhenjiang Jiangsu 212013,China）
Abstract：Objective: To construct and identify the expression vector expressing the protein of modified glucagon-like peptide-1(mGLP-1) containing His-tag in Bacillus subtilis WB800N. Methods: The mGLP-1 gene including of His-tag and BamH Ⅰ/Xba Ⅰ restriction enzyme cutting sites was obtained by chemical synthesis, and then inserted into the Escherichia coli-Bacillus subtilis shuttle vector pHT43 carrying the signal peptide amyQ sequence to construct the recombinant plasmid pHT43-mGLP-1. The plasmid pHT43-mGLP-1 was transformed into Bacillus subtilis WB800N by electroporation. Colony PCR, sequencing technologies, Tricine SDS-PAGE and Western blotting were used to identify the recombinant Bacillus subtilis WB800N/pHT43-GLP-1. Results: The result of colony PCR and sequencing showed that the recombinant plasmid pHT43-mGLP-1 was successfully transformed into Bacillus subtilis WB800N. Tricine SDSPAGE and Western blotting showed the transformed bacterium successfully expressed and released the target protein mGLP-1 to culture supernatant, and the protein expression was significantly increased by the induction of 1 mmol/L IPTG. Conclusion: The recombinant expression vector carrying mGLP-1 was successfully constructed and target protein could be expressed in Bacillus subtilis WB800N, which will be benefical for further explore the animal experiments and micro-organism preparations.
［Key words］glucagon-like peptide-1; Bacillus subtilis; expression vector
2019, Vol. 29 
