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巴马小型猪内源性反转录病毒的特性
Characterization of Porcine Endogenous Retrovirus in BaMa Miniature Pig
【作者】 斯日古楞;
【作者基本信息】 内蒙古农业大学 , 基础兽医学, 2003, 硕士
【摘要】 本实验结合国内开展的相关科研项目进行了猪内源性反转录病毒(porcine endogenous retrovirus,PERV)的前期研究工作,现分述如下: 1.巴马小型猪中内源性反转录病毒的存在与表达。应用PCR和RT-PCR的方法,对来源于巴马小型猪外周血淋巴细胞(PBL)的DNA和RNA样品进行了系统的检测。结果表明,在所有被检样品中均有PERV前病毒的存在,且存在于基因组中的前病毒均能以mRNA的形式进行表达。 2.PERV体外感染人HEK293细胞模型的建立。应用猪淋巴细胞和G418抗性293细胞共培养、经G418加压筛选的方法成功地建立了PERV感染HEK293细胞的模型。这一感染模型的成功建立,为今后的研究工作提供了一个极好的实验平台。 3.巴马小型猪来源的PERV囊膜基因的克隆与序列测序。应用RT-PCR的方法从巴马小型猪外周血淋巴细胞RNA样品中钓取了PERV-env基因,并克隆至pGEM-T Easy载体。序列测定表明,该克隆片段由2013bp组成,编码671个氨基酸。 综上所述,本课题的研究工作已具有良好基础,为进一步深入研究PERV创造了条件。
【Abstract】 The porcine endogenous retro viruses (PERV) was studied in the research, which is relevant to science research project at home and abroad. The research is divided into three parts:The existence and expression of PERV from BM miniature pig. We detected DNA and RNA samples of the peripheral blood lymphocyte cell from BM miniature pig. The reswlt showed that PERV proviruses is existed in all tested samples, and the virus are expressed by mRNA.A model of the HEK293cell infected by PERV in vitro is obtained. It is important for us to further research.We clon,ed and analysized the envelope gene of PERV. We obtained PERV-env gene from the peripheral blood lymphocyte cell RNA samples of BM miniature pig using RT-PCR, then these gene was cloned into pGEM-T Easy vectors. Sequence analysis shows that is made up of 2013 base pair and coding 671 amino acids.In view of what I have mentioned above,the research is facility to PERV for further research.
【Key words】 porcin endogenous retrovirus; BM miniature pig; detection; cell model; colon;
- 【网络出版投稿人】 内蒙古农业大学 【网络出版年期】2003年 03期
- 【分类号】S858.28
- 【下载频次】89