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猪白介素18基因在BHK细胞中的表达

Expression of porcine interleukin18 in the BHK cell
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摘要 将猪白介素18(Porcine interleukin18,PIL-18)基因亚克隆到真核表达载体pEGFP-N1中,构建了重组表达质粒pEGFP-PIL-18,利用脂质体法转染BHK细胞,采用荧光显微镜实时观察和RT-PCR检测技术确定目的蛋白的表达情况.结果在转染重组质粒24 h4、8 h后的BHK细胞中均观察到绿色荧光,转染的BHK细胞经G418筛选14 d后,用RT-PCR法检测目的基因,并扩增到长576 bpd的目的cDNA片段。表明在BHK细胞中成功地表达了PIL-18与EGFP的融合蛋白,这为下一步建立稳定表达PIL-18的细胞系奠定了基础. Porcine IL-18 gene was subcloned into the eukaryotic expression vector pEGFP--N1 to construct the recombinant plasmid pEGFP-PIL-18, which was transfected into the BHK cell by using Lipofectin. The expression of PIL--18 was determined by fluoroscopy and RT-PCR, 24 hours and 48 hours after transfecting,the Green fluorescence can be observed under fluorescence microscope and a 576 bp cDNA was detected by RT--PCR 14 days after screening with G418. The results indicated that fusion protein of PoIL--18 and EGFP was successfully expressed in BHK cell. This method can be further used to establish a cell line stably expressing PIL-18.
作者 王笑笑 宗瑞谦 吕转萍 窦永喜 宋世斌 景志忠
机构地区 甘肃农业大学动物医学院 巴音郭棱职业技术学院生物工程系 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室甘肃省动物寄生虫病重点实验室
出处 《甘肃农业大学学报》 CAS CSCD 2007年第6期23-27,共5页 Journal of Gansu Agricultural University
基金 兰州市科技攻关项目(05-1-47).
关键词 猪白介素18 转染 BHK细胞 真核表达 porcine interleukin18 transfection BHK cell eukaryotic expression
分类号 S852.42 [农业科学—基础兽医学] Q785 [生物学—分子生物学]
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参考文献14

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甘肃农业大学学报
2007年 第6期

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