甘蔗花叶病毒CP基因的高效表达和抗血清制备

High-level expression and antiserum preparation of CP gene of sugarcane mosaic virus

将甘蔗花叶病毒的CP基因克隆到表达载体pET22b(+),并在大肠杆菌BL21(DE3)中得到高效表达,表达蛋白约占总蛋白的15%。用此蛋白制备了效价高专化性强的抗体。此方法可以解决制备马铃薯Y病毒属病毒的血清时遇到的病毒提纯产量低和寄主蛋白影响等问题。

The coat protein gene of sugarcane mosaic virus(SCMV)was cloned into expression vector pET22b(+),and transferred into E.coli BL21(DE3).It can be expressed as a fusion protein to high level(about 15%of the total protein)when induced with IPTG.The molecular weight of the fusion protein is ca.38kD.The results of Western blotting confirmed the fact that the SCMV was expressed correctly.With the protein expressed in E.coli,antiserum were prepared with high titer and specificity.