| 肠道病毒71型P1和3CD基因双顺反子重组腺病毒的构建及表达 |
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| 引用本文： | 戈小琴,李鸿钧,王文举,谢天宏,张光明,孙茂盛. 肠道病毒71型P1和3CD基因双顺反子重组腺病毒的构建及表达[J]. 中国生物制品学杂志, 2010, 23(4) |
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| 作者姓名： | 戈小琴 李鸿钧 王文举 谢天宏 张光明 孙茂盛 |
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| 作者单位： | 中国医学科学院,北京协和医学院医学生物学研究所,昆明,650118 |
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| 摘 要： | 目的构建肠道病毒71型(EV71)P1和3CD基因双顺反子重组腺病毒,并检测3CD蛋白酶的切割作用和表达产物的抗原性。方法采用RT-PCR法从阜阳分离的EV71中扩增P1和3CD基因,构建穿梭质粒pShuttle-CMV-P1-3CD、pShuttle-CMV-P1和pShuttle-CMV-3CD;经同源重组获得重组腺病毒rAd-P1-3CD、rAd-P1和rAd-3CD,分别转染293细胞,RT-PCR法检测目的基因的转录,免疫荧光法检测目的蛋白的表达。结果3种重组腺病毒经PCR和酶切鉴定表明构建正确;RT-PCR检测表明,3个重组腺病毒均已转录目的基因mRNA;免疫荧光检测仅观察到rAd-P1-3CD表达产物具有特异性,而rAd-P1、rAd-3CD未能检测到特异性表达产物。结论已成功构建EV71P1和3CD基因双顺反子重组腺病毒,表达产物具有EV71特异抗原性,提示P1产物只有在被3CD蛋白酶切割后才体现抗原性。
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| 关 键 词： | 肠道病毒71型 P1基因 3CD基因 双顺反子 重组腺病毒 |
Construction and Expression of Bicistronic Recombinant Adenovirus with P1 and 3CD Genes of Enterovirus 71 |
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| GE Xiao-qin,LI Hong-jun,WANG Wen-ju,et al. Construction and Expression of Bicistronic Recombinant Adenovirus with P1 and 3CD Genes of Enterovirus 71[J]. Chinese Journal of Bilogicals, 2010, 23(4) |
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| Authors: | GE Xiao-qin LI Hong-jun WANG Wen-ju et al |
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| Affiliation: | GE Xiao-qin,LI Hong-jun,WANG Wen-ju,et al(Institute of Medical Biology,Academy of Medical Sciences , Peking Union Medical College,Kunming 650118,China) |
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| Abstract: | Objective To construct a bicistronic recombinant adenovirus with P1 and 3CD genes of enterovirus 71(EV71) and determine the cleavage effect of 3CD protease as well as the antigenicity of expressed product. Methods P1 and 3CD genes were amplified from EV71 strain isolated from Fuyang City, Anhui Province, China for construction of shuttle plasmids pShuttle-CMVP1-3CD, pShuttle-CMV-P1 and pShuttle-CMV-3CD, based on which recombinant adenoviruses rAd-P1-3CD, rAd-P1 and rAd-3CD were obtained by homologous recomb... |
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| Keywords: | Enterovirus 71(EV71) P1 gene 3CD gene Bicistronic Recombinant adenovirus |
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