日本血吸虫Ⅴ型胶原蛋白α1链的原核表达分析

EXPRESSION OF SCHISTOSOMA JAPONICUM TYPE V COLLAGEN A1 CHAIN IN E.COLI BL21

目前广泛应用的各类胶原及胶原蛋白多为人工提取的方法获得,为了得到高纯度原核表达的胶原蛋白,本研究构建了血吸虫V型胶原蛋白(Schistosoma japonicum type V collagen,Sj Col V)(a1)基因AY815998.1的原核表达质粒,并对其进行了原核表达。结果表明SjColV(a1)蛋白能在大肠杆菌BL21(DE3)菌株中高效表达,但原核表达的融合蛋白不够稳定,且存在形式多样。本文就其表达情况进行了分析,并纯化得到原核表达的全长rSjColV(a1),用V型胶原蛋白的单抗对其进行鉴定,同时分析其在?80℃保存的稳定性,研究结果为SjColV(a1)的深入研究和利用提供了一定的理论依据。

To analyze expression of Schistosoma japonicum type V collagen a1 chain in E.coli BL21（DE3）,a prokaryotic expression vector pET28-SjColV（a1） was constructed.rSjColV（a1） was expressed in E.coli BL21（DE3） with induction of IPTG and confirmed in SDS-PAGE and Western blot.The results showed that SjColV（a1） was efficiently expressed in BL21（DE3）,but the products were unstable.In this study we analyzed conditions for prokaryotic expression of SjCol V（a1） and proposed future studies for the application of SjColV（a1） protein.