摘要
目的:克隆丙型肝炎病毒核心蛋白基因及其上游DNA序列,为此基因的表达研究作准备。方法:用反转录和PCR方法从HCV的总RNA中扩增得到核心蛋白基因及其上游DNA序列,连接到pMD18-T载体上,用限制性内切酶切下目的基因,插入到巴斯德毕赤酵母表达载体pPIC9K中,构建成重组质粒,测序证明正确后,再将目的基因在毕赤酵母中进行克隆,鉴定。结果:重组质粒转化毕赤酵母后,经PCR鉴定,证明形成了目的基因的克隆。结论:应用毕赤酵母作为受体菌,pPIC9K为载体,成功克隆了HCV核心蛋白基因。
Objective： To clone the HCV core protein gene and its upstream DNA for the expression of the gene. Methods： The core protein gene and its upstream DNA were got from HCV total RNA by reverse transcription and PCR.They were linked into pMD18 - T vector and were cut down by restriction endonuclease. Then they were inserted into expression vector pPIC9K of Pichia pastoris, and the recombinant plasmids were obtained.After they were identified by sequence , the target gene was cloned and identified. Results： After Pichia pastoris was transformed by the recombinant plasmid, the cloned target gene was confirmed by PCR analysis. Conclusion： The HCV core protein gene was cloned successfully by using pPIC9K as the vector and using Pichia pastoris as host bateria.
出处
《生物技术》
CAS
CSCD
2007年第2期1-3,共3页
Biotechnology
基金
山东省自然基金项目资助("辣根过氧化物酶与丙型肝炎病毒核心蛋白的融合表达研究"
编号:Y2004C33)
