人巨细胞病毒pp65基因片段毕赤酵母表达载体的构建

目的 构建人巨细胞病毒pp65基因片段毕赤酵母表达载体.  方法   PCR扩增pp651087-1515 nt,克隆于pMD19-T载体,转化大肠杆菌DH5α.提取质粒测序鉴定后,与pPIC9K载体连接,转化DH5α,筛选阳性克隆,酶切及测序鉴定.   结果  获得了重组酵母表达载体pPIC9K-pp65,测序结果证实为pp65 1087-1515 nt基因,与基因库报道序列一致.  结论 构建得到人巨细胞病毒pp651087-1515 nt毕赤酵母表达载体.

Construction of expression vector of Pichia pastoris for human cytomegalovtrus pp65 gene fragment

Objective To construct Pichia expression vector carrying gene coding for human cytomegalovirus (HCMV) pp65. Methods The HCMV pp65 1087-1515 nt gene was amplified with PCR and cloned into pMD19-T vector.The recombinant plasmid was transformed into E.coli DH5α.The positive clone was screened and identified by sequence analysis.The enzyme-restricted fragment was subcloned into pPIC9K, and the recombinant product was transformed into DH5a and identified by restriction enzyme digestion and sequence analysis. Results The yeast eukaryotic expression vector pPICgK-pp65 was constructed and itS sequence was the same as that reported in Genebank. Conclusion The Pichia expression vector carrying HCMV pp65 1087-1515 nt can be constructed.