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中国农学通报 ›› 2012, Vol. 28 ›› Issue (20): 88-92.doi: 10.11924/j.issn.1000-6850.2012-0213

• 兽医—应用研究 • 上一篇    下一篇

犬SLAM基因真核表达载体的构建及Vero细胞系转染的研究

苏建青 褚秀玲 张吉清 马秀亮 江成   

  • 收稿日期:2012-01-29 修回日期:2012-03-07 出版日期:2012-07-15 发布日期:2012-07-15
  • 基金资助:

    表达犬瘟热病毒细胞受体信号淋巴细胞激活因子细胞系的建立

Construction of Eukaryotic Expressing Vector of SLAM Gene and its Transient Expression in Vero Cell

  • Received:2012-01-29 Revised:2012-03-07 Online:2012-07-15 Published:2012-07-15

摘要:

为了研究犬瘟热病毒细胞受体SLAM基因的特性和功能。将基因SLAM克隆到真核表达载体pcDNA3.1(+)上,构建真核表达载体pCDNA3.1/SLAM,重组质粒纯化后应用脂质体2000转染到Vero细胞中,通过RT-PCR和免疫印迹方法检测犬SLAM基因在Vero细胞中的转录和表达情况。结果表明:成功构建了表达SLAM基因的真核表达载体,RT-PCR和免疫印迹显示SLAM基因获得表达;pcDNA3.1/SLAM载体的构建为研究犬SLAM基因在Vero细胞中的稳定表达奠定了基础。

关键词: 抗性评价, 抗性评价

Abstract:

To elucidate the character and functions of cellular receptors of canine distemper virus, the gene SLAM of canine distemper virus’s open reading frame (ORF) was cloned into eukaryotic expression vector pCDNA3.1(+) to generate the recombinant plasmid pcDNA3.1/SLAM. The pureed plasmid was transected into Vero cells in vitro with Lipofectamine 2000. The transient expression of the SLAM protein was detected by reverse transcription polymerase chain reaction and Western-blot assay. The results showed that the eukaryotic expression vectors of canine SLAM gene were constructed successfully. The reverse transcription polymerase chain reaction and Western-blot assay confirmed that the protein SLAM was expression in Vero cell. Recombinant SLAM was successfully expressed, which laid foundation for further research on the stable express of SLAM gene in Vero.