| 犬新孢子虫NcSRS2基因真核表达质粒的构建 |
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| 引用本文： | 赵占中,刘群,汪明.犬新孢子虫NcSRS2基因真核表达质粒的构建[J].畜牧兽医学报,2005,36(8):819-822. |
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| 作者姓名： | 赵占中 刘群 汪明 |
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| 作者单位： | 中国农业大学动物医学院,北京,100094 |
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| 基金项目： | 国家自然基金资助项目（30371080）;北京市自然科学基金资助项目（6042016） |
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| 摘 要： | 根据犬新孢子虫NcSRS2基因序列，设计了1对含有Kozak序列、PstⅠ和XbaⅠ酶切位点的引物，以含有NcSRS2基因的质粒P43为模板，经PCR扩增获得NcSRS2 ORF基因片段，用PstⅠ和XbaⅠ双酶切该片段，回收得到含有以上2个酶切位点黏端的NcSR2 ORF基因，将此基因片段克隆至相同酶切回收后的pcDNA3．1（＋）真核表达载体中，获得重组质粒pcNCSRS2。经PCR鉴定、限制性内切酶分析和克隆片段序列测定、比较，证实了重组质粒的正确性。
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| 关 键 词： | 犬新孢子虫 聚合酶链反应 NcSRS2基因 真核表达载体pcDNA3．1（＋） |
| 文章编号： | 0366-6964（2005）08-0819-04 |
| 收稿时间： | 2004-05-19 |
| 修稿时间： | 2004-05-19 |
Construction of an Eukaryotic Expression Plasmid for NcSRS2 Gene of Neospora caninum |
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| ZHAO Zhan-zhong,LIU Qun,WANG Ming.Construction of an Eukaryotic Expression Plasmid for NcSRS2 Gene of Neospora caninum[J].Acta Veterinaria et Zootechnica Sinica,2005,36(8):819-822. |
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| Authors: | ZHAO Zhan-zhong LIU Qun WANG Ming |
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| Abstract: | Based on the NcSRS2 gene sequence of Neospora caninum, a pair of primers containing Kozak sequence, Pstand Xbaenzyme digestion sites were designed .Using the plasmid P43 which contains NcSRS2 gene as template,the ORF region of NcSRS2 gene was amplified by polymerase chain reaction (PCR), then the PCR product was digested with Pstand Xba,and the eukaryotic expression vector pcDNA3.1(+)was also done ,then the gene was cloned into the vector.Finally a recombinant plasmid named pcNcSRS2 was obtained, and was identified by PCR, restriction enzyme analysis and sequencing. |
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| Keywords: | Neosproa caninum PCR NcSRS2 gene eukaryotic expression vector pcDNA3 1(+) |
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