期刊论文

中文

安徽农业科学

0517-6611

2012

13

7739-7741+7803

10.13989/j.cnki.0517-6611.2012.13.142

国家科技重大专项项目（2012ZX10004220-008）； 中央级公益性科研院所基本科研业务费专项资金项目（2010JB12,2012JB16）； 上海市科技兴农重点攻关项目[沪农科攻字（2005）第3-4号]；

本校为第一机构

[目的]构建微小隐孢子虫的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pVAXⅠ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的功能奠定了基础。

[ Objective ] This study aimed to construct and preliminarily identify the eukaryotic expression vector of C. 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 identified 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 expre...