| H7亚型禽流感病毒HA1基因的克隆与原核表达 |
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| 引用本文： | 庞平,唐懿,雷霆,武山,李强,陈晨,曹红,陈福勇.H7亚型禽流感病毒HA1基因的克隆与原核表达[J].中国家禽,2006,28(23):10-13. |
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| 作者姓名： | 庞平 唐懿 雷霆 武山 李强 陈晨 曹红 陈福勇 |
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| 作者单位： | 中国农业大学动物医学院,北京,100094 |
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| 摘 要： | 利用PCR技术亚克隆了H7亚型禽流感病毒的HA1基因,并将PCR产物连接到克隆载体pMD 18-T Simple vector,转化大肠杆菌。测序结果表明所克隆的片断大小为1035bp。将HA1基因克隆至原核表达载体pGEX-6P-1,经酶切和PCR鉴定,证明成功构建了重组表达载体pGEX-HA1。将构建好的融合表达载体在IPTG的诱导下在大肠杆菌中得到了表达。融合蛋白GST-HA1的分子量为63ku。Western-Blot和ELISA鉴定结果表明,融合蛋白与H7亚型禽流感阳性血清发生特异性反应,而与H5、H9亚型抗血清不发生反应。
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| 关 键 词： | 禽流感病毒 HA1基因 基因克隆 表达 抗原性 |
| 收稿时间： | 2006-07-29 |
| 修稿时间： | 2006年7月29日 |
Cloning and Prokaryotic Expression of HA1 Gene of H7 Avian Influenza Virus |
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| Pang Ping,Tang Yi,Lei Ting,Wu Shan,Li Qiang,Chen Chen,Cao Hong,Chen Fuyong.Cloning and Prokaryotic Expression of HA1 Gene of H7 Avian Influenza Virus[J].China Poultry,2006,28(23):10-13. |
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| Authors: | Pang Ping Tang Yi Lei Ting Wu Shan Li Qiang Chen Chen Cao Hong Chen Fuyong |
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| Institution: | College of Veterinary Medicine,China Agricultural University,Beijing 100094 |
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| Abstract: | In this study,HA1 gene was subcloned by PCR,ligated into pMD18-T simple vector and then transformed E.coli. Gene sequencing analysis showed that the cloned HA1 was 1 035 bp in length. The HA1 gene was inserted into the expression plasmid pGEX-6P-1. The recombinant fusion protein was expressed in E.coli BL21 induced by IPTG in the form of inclusion bodies. The molecular weights of GST-HA1 is 63 ku as demonstrated by western-blotting using AIV antibody against H7 subtype. The result of ELISA developed with this recombinant protein showed that the recombinant protein can only react with antisera of H7 subtype. |
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| Keywords: | avian influenza virus HA1 gene gene clone gene expression antigenicity |
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