| 斯氏艾美尔球虫肌动蛋白解聚因子在HeLa细胞中的表达 |
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| 引用本文： | 任保彦,付成福,宫鹏涛,韩乾忠,杨举,李赫,张国才,张西臣,李建华.斯氏艾美尔球虫肌动蛋白解聚因子在HeLa细胞中的表达[J].中国预防兽医学报,2011,33(12). |
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| 作者姓名： | 任保彦 付成福 宫鹏涛 韩乾忠 杨举 李赫 张国才 张西臣 李建华 |
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| 作者单位： | 吉林大学畜牧兽医学院,吉林 长春,130062 |
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| 基金项目： | 国家863计划，新世纪优秀人才支持计划资助项目 |
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| 摘 要： | 为真核表达斯氏艾美尔球虫(Eimeria stiedae)肌动蛋白解聚因子(ADF)基因,本研究采用RT-PCR技术扩增ADF基因,构建pVAX- 1-ADF真核重组质粒,并转染HeLa细胞.采用RT-PCR方法检测ADF mRNA转录水平、western blot检测ADF蛋白表达水平.琼脂糖凝胶电泳结果表明PCR扩增出一条大小为350 bp的特异性片段,与预期值相符.Western blot检测显示,转染pVAX- 1-ADF的HeLa细胞在大约13ku处出现特异性表达条带,与预期值相符.本研究构建了E.stiedae ADF基因真核表达质粒pVAX- 1-ADF,并且在体外真核细胞中表达,为进一步寻找防治E.stiedae的疫苗候选基因奠定了基础.
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| 关 键 词： | 斯氏艾美尔球虫 ADF基因 真核表达 |
Expression of Eimeria stiedae actin depolymerizing factor in HeLa cells |
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| REN Bao-yan,FU Cheng-fu,GONG Peng-tao,HANQian-zhong,YANG Ju,LI He,ZHANG Guo-cai,ZHANG Xi-chen,LI Jian-hua.Expression of Eimeria stiedae actin depolymerizing factor in HeLa cells[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(12). |
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| Authors: | REN Bao-yan FU Cheng-fu GONG Peng-tao HANQian-zhong YANG Ju LI He ZHANG Guo-cai ZHANG Xi-chen LI Jian-hua |
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| Institution: | REN Bao-yan,FU Cheng-fu,GONG Peng-tao,HAN Qian-zhong,YANG Ju,LI He,ZHANG Guo-cai,ZHANG Xi-chen,LI Jian-hua(College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China) |
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| Abstract: | To express the actin depolymerizing factor(ADF) of Eimeria stiedae in HeLa cells,the ADF gene was amplification from the E.stiedae total RNA by RT-PCR and cloned into pVAX-1.The resultant recombinant plasmid was transfected into HeLa cells with FuGENE.The results indicated that the ADF gene of E.stiedae was expressed in HeLa cells detected by RT-PCR and western blot,which would be useful for evaluating immunogenicity of ADF gene as a DNA vaccine against E.stiedae. |
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| Keywords: | Eimeria stiedae actin depolymerizing factor gene eukaryotic expression |
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