旋毛虫表皮胶原蛋白基因的原核表达

目的构建表皮胶原蛋白基因X21的原核表达载体pET28-X21ex,诱导其表达胶原蛋白。方法应用PCR技术扩增目的基因X21,纯化后将其克隆入表达载体pET28a,重组质粒经酶切鉴定后,转入大肠杆菌DE3诱导表达,利用SDS-PAGE检测胶原蛋白的表达效果。结果利用DNA重组技术构建了表皮胶原蛋白基因X21的原核表达载体pET28-X21ex,转化大肠杆菌后经诱导表达了30kDa的融合蛋白,以诱导3h的表达量最高,在35%以上。结论旋毛虫表皮胶原蛋白基因X21在大肠杆菌中能够高效表达,为进一步研究旋毛虫表皮胶原蛋白的结构与功能奠定了基础。

Prokaryotic Expression of Trichinella spiralis Cuticle Collagen Gene

Objective To construct the prokaryotic expression vector pET28-X21ex of cuticle collagen gene X21and induce it to express collagen.Methods PCR technique was used to amplify the target gene X21.After purifying the gene was cloned into expression vector pET28a.The recombinant plasmid was transformed into E.coli DE 3 and induced after identified with restriction enzyme digestion.The expression level of collagen was tested by SDS-PAGE.Results Using the recombinant DNA technique the prokaryotic expression vector pET28-X21ex of cuticle collagen gene was constructed successfully.After transforming into E.coli and inducing the30kDa fusion protein was expressed.The level of expression peaked at3h post-incubation,which was more than35%of all the proteins.Conclusion The high level expression of trichinella spiralis cuticle collagen gene X21was obtained in E.coli which will give the base of researching the constructions and functions of Trichinella spiralis cuticle collagen.