| 猪白细胞介素-2在BHK-21细胞中表达及其活性分析 |
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| 引用本文： | 王大伟,乔军,张辉,胡圣伟,丁波,陈创夫.猪白细胞介素-2在BHK-21细胞中表达及其活性分析[J].中国预防兽医学报,2011,33(5). |
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| 作者姓名： | 王大伟 乔军 张辉 胡圣伟 丁波 陈创夫 |
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| 作者单位： | 石河子大学动物科技学院,新疆,石河子,832000 |
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| 基金项目： | 转基因生物新品种培育国家重大专项 |
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| 摘 要： | 为鉴定BHK-21细胞中稳定表达猪白细胞介素-2(PoIL-2)的活性,本研究利用RT-PCR技术从猪的淋巴细胞中克隆PoIL-2基因,将其连接于pcDNA3.1载体中,并转染BHK-21细胞.经过G418筛选及RT-PCR鉴定,获得稳定表达PoIL-2基因的BHK-21细胞系.采用实时定量PCR及ELISA测定PoIL-2在BHK-21细胞内的表达,同时以淋巴细胞增殖试验(MTT)检测PoIL-2蛋白活性.试验结果表明稳定整合PoIL-2基因的BHK-21细胞系其PoIL-2 mRNA的转录效率是阴性对照组1.43倍,蛋白表达量是阴性对照组1.29倍.MTT试验表明真核表达的PoIL-2促淋巴细胞增殖活性与阴性对照组相比差异极显著(p＜0.01),表明BHK-21细胞表达的PoIL-2具有良好的生物学活性.本研究为开发新型基因工程疫苗佐剂和抗病毒制剂奠定了基础.
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| 关 键 词： | 猪白细胞介素2 真核表达 实时定量PCR 淋巴细胞增殖试验 |
Expression of the porcine interleukin-2 in BHK-21 cells and bioactivity analysis |
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| WANG Da-wei,QIAO Jun,ZHANG Hui,HU Sheng-wei,DING Bo,CHEN Chuang-fu.Expression of the porcine interleukin-2 in BHK-21 cells and bioactivity analysis[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(5). |
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| Authors: | WANG Da-wei QIAO Jun ZHANG Hui HU Sheng-wei DING Bo CHEN Chuang-fu |
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| Institution: | WANG Da-wei,QIAO Jun,ZHANG Hui,HU Sheng-wei,DING Bo,CHEN Chuang-fu (College of Animal Science & Technology,Shihezi University,Shihezi 832000,China) |
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| Abstract: | To express porcine interleukin-2 (PoIL-2) in eukaryotic cells and evaluate its biological activity, the PoIL-2 gene was amplified from pig lymphocyte by RT-PCR and inserted into pcDNA3.1 vector, which was ultimately transfected into BHK-21 cells. A BHK-21 cell line expressing PoIL-2 gene was established by G418 screen and RT-PCR identification. Gene expression level and activity of PoIL-2 in BHK-21 cells was examined by Real-time PCR, ELISA, and lymphocyte proliferation test (MTT), respectively. The result ... |
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| Keywords: | porcine interleukin-2 eukaryotic expression real-time PCR MTT |
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