BVDV P7蛋白的原核表达及其生物信息学分析

Prokaryotic Expression and Bioinformatics Analysis of BVDV P7 Protein

为了对BVDV-1 P7蛋白进行原核表达及生物信息学分析,采用RT-PCR的方法扩增BVDV NADL株BVDV-1 P7的ORF,扩增产物经Bam H I、Sal I双酶切后插入原核表达载体p EGX4T-1(+),构建P7基因原核表达载体并将其转化至工程菌E.coli BL21(DE3)中进行诱导表达,并对其进行生物信息学预测。结果表明,成功扩增并构建了BVDV NADL株的BVDV-1 P7原核表达载体,命名p EGX4T-1-P7。SDS-PAGE和Western blot表明,在分子质量34 k Da的位置出现与预期大小一致的蛋白条带;生物信息学结果表明在P7蛋白有信号肽,存在疏水区。该研究成功的表达了P7蛋白,该蛋白是一个疏水性蛋白,为其进一步的功能研究奠定了坚实的基础。

For prokaryotic expression of BVDV P7 gene and its bioinformatics analysis, RT-PCR method was used to amplify BVDV P7 gene which isolated from BVDV-1 NADL,then BamH I and Sol I were used to digest PCR products and this recovered fragment was inserted into the pEGX4T-1（＋）, a prokaryotic expression vector,to generate recombinant pEGX4T-1-P7.pEGX4T-1-P7 was transformed into E.coli B1221 ,using bioinformatics technology for bioinformatics prediction at the same time.The results showed that amplification was successful and the BVDV NADL strains of BVDV-1 P7 prokaryotic expression vector were built,named pEGX4T- 1-P7.SDS-PAGE and Western blot showed that at expected size a molecular mass of 34 kDa appeared the expressed protein.The bioinformatics analysis by prediction software indicated that the signal peptide and hydrophobic site existed in P7 protein.h concluded that P7 protein was successfully expressed and it was a hydrophobic protein,which established solid foundation for the further study of its functions.