CAJ | 학술논문

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.
목적 구건인거세포병독pp65기인편단필적효모표체재체.방법 PCR확증pp651087-1515 nt,극륭우pMD19-T재체,전화대장간균DH5α.제취질립측서감정후,여pPIC9K재체련접,전화DH5α,사선양성극륭,매절급측서감정.결과 획득료중조효모표체재체pPIC9K-pp65,측서결과증실위pp65 1087-1515 nt기인,여기인고보도서렬일치.결론 구건득도인거세포병독pp651087-1515 nt필적효모표체재체.
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.