丝心蛋白基因分子克隆与表达的初步探讨

A Preliminary Study on Expression of Fibroin C Subunit as a Fusion Protein in E. Coli

通过聚合酶链反应（ＰＣＲ）扩增丝心蛋白Ｃ亚基结构基因，并将其克隆到融合蛋白表达载体ｐＲＩＴ２Ｔ质粒中得到ｐＲＩＴ２Ｔ－ＦＬ质粒，在大肠杆菌株Ｐ２３９２内进行表达．十二烷基硫酸钠聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）和免疫印迹反应证明融合蛋白在大肠杆菌中得到了表达．

The structural gene of fibroin C subunit(i.e. fibroin light chain, FL) was amplified by polymerase chain reaction(PCR) from pFL18 plasmid, which carries a cDNA clone of fibroin C subunit. The PCR product was cloned on fusion protein expression vector pRIT2T, and was expressed in the E. Coli strain P2392. SDS polyacrylamide gel electrophoresis(SDS PAGE) and immunoblotting were carried out to verify the expression of protein A fibroin C subunit fusion protein.