Construction and Preliminary Identification of Eukaryotic Expression Vector of Cryptosporidium parvum miR-2980  

微小隐孢子虫miR-2980真核表达载体的构建及初步鉴定(英文)

作　　者：呼高伟[1,2] 程天印[1] 米荣升[2] 秦培兰[2] 黄燕[2] 周鹏[2] 曹薇[2] 陈兆国[2] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]中国农业科学院上海兽医研究所,农业部动物寄生虫学重点开放实验室,中国农业科学院动物源性食品安全研究中心,上海200241

出　　处：《Agricultural Science & Technology》2012年第5期1093-1096,共4页农业科学与技术（英文版）

基　　金：Supported by National Major Special Science and Technology Project of China(2012ZX10004220-008);Basic Scientific Research Operational Fund for Central-level Public-interest Research Institutes (2010JB12,2012JB16);Key Project of Science and Technology to Develop Agriculture in Shanghai (2005 No. 3-4)~~

摘　　要：[Objective] This study aimed to construct and preliminarily identify the eu- karyotic expression vector of Cryptosporidium parvum miR-2980. [Method] The cp-miR- 2980 precursor was amplified from C. parvum genomic DNA and cloned into pMD18- T vector. The amplified precursor was then subcloned into pVAX I vector and identi- fied with restriction endonuclease digestion and sequencing. The recombinant plasmid pVAX-miR2980 was transfected into HCT-8 cells. Total RNA was extracted and the expression of cp-miR-2980 was evaluated by RT-PCR detection. [Result] The results showed that the recombinant eukaryotic expression vector pVAX-miR2980 was suc- cessfully constructed, which can express cp-miR-2980 in HCT-8 cell. [Conclusion] This study laid the foundation for further exploring the biological function of cp-miR-2980.[目的]构建微小隐孢子虫miR-2980真核表达载体,并进行鉴定。[方法]从微小隐孢子虫(Cryptosporidium parvum)基因组DNA扩增cp-miR-2980前体,克隆到pMD18-T载体上,经测序鉴定正确后亚克隆到pVAX1载体上,转染HCT-8细胞,提取细胞总RNA,RT-PCR检测cp-miR-2980的表达。[结果]成功地构建了cp-miR-2980真核表达载体pVAX-miR2980,RT-PCR检测证实其能在真核细胞HCT-8中表达。[结论]构建的真核表达载体pVAX-miR2980能在HCT-8细胞中成功表达cp-miR-2980,为后续研究cp-miR-2980的功能打下了基础。

关 键 词：cp-miR-2980 PRECURSOR Eukaryotic expression vector RT-PCR 

分 类 号：S852.723[农业科学—基础兽医学]