| 微小隐孢子虫miR-2980真核表达载体的构建及初步鉴定 |
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| 引用本文： | 呼高伟,程天印,米荣升,秦培兰,黄燕,周鹏,曹薇,陈兆国.微小隐孢子虫miR-2980真核表达载体的构建及初步鉴定[J].安徽农业科学,2012(13):7739-7741,7803. |
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| 作者姓名： | 呼高伟 程天印 米荣升 秦培兰 黄燕 周鹏 曹薇 陈兆国 |
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| 作者单位： | 湖南农业大学动物医学院,湖南长沙410128;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;湖南农业大学动物医学院,湖南长沙,410128;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 |
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| 基金项目： | 国家科技重大专项项目，中央级公益性科研院所基本科研业务费专项资金项目，上海市科技兴农重点攻关项目 |
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| 摘 要： | 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。
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| 关 键 词： | cp-miR-2980 前体 真核表达载体 RT-PCR |
Construction and Preliminary Identification of Eukaryotic Expression Vector of Cryptosporidium parvum miR-2980 |
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| Institution: | HU Gao-wei et al(College of Veterinary Medicine,Hunan Agricultural University,Changsha,Hunan 410128) |
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| Abstract: | 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 pVAXⅠvector 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 expression vector pVAX-miR2980 was successfully 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. |
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| Keywords: | cp-miR-2980 Precursor Eukaryotic expression vector RT-PCR |
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