A Method of Construction Polycistron Tandem Gene of Small Peptide / 中国生物工程杂志

ABSTRACT

Objective: To construct a polycistron tandem repeated Echistatin (Ecs) gene. Methods: Three Ecs genes with independent initiation and termination codon were ligated tandem through restriction enzyme sites after amplified with 3 pairs of primers using pMD18T-Ecs as template. The polycistron Ecs gene was inserted into pET30a and expressed in E.coli BL21(DE3) with IPTG induction. The expression results were identified by 18% SDS-PAGE and Western blot. Results: The expression of Ecs polycistron was accomplished with 18% expression level of total protein determined by SDS-PAGE and Western blot. Conclusion: The successful expression of Ecs polycistron provided a new method for the preparation of low molecular weight protein.