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| H5N1亚型禽流感病毒NS1基因的克隆及在E.coli中的表达 | |
| 引用本文： | 刘威龙,焦培荣,刘畅,阿合买提·买买提,陈化兰.H5N1亚型禽流感病毒NS1基因的克隆及在E.coli中的表达[J].新疆农业大学学报,2007,30(3):69-72. |
| 作者姓名： | 刘威龙 焦培荣 刘畅 阿合买提·买买提 陈化兰 |
| 作者单位： | 1. 新疆农业大学,动物医学学院,乌鲁木齐,830052;中国农业科学院哈尔滨兽医研究所,农业部动物流感重点开放实验室,哈尔滨,150001 2. 中国农业科学院哈尔滨兽医研究所,农业部动物流感重点开放实验室,哈尔滨,150001 3. 中国农业科学院哈尔滨兽医研究所,农业部动物流感重点开放实验室,哈尔滨,150001;内蒙古农业大学,动物科学与医学学院,呼和浩特,010018 4. 新疆农业大学,动物医学学院,乌鲁木齐,830052 |
| 摘 要： | 选择一株H5N1亚型高致病性禽流感病毒株A/Goose/Guangdong/1/96(简写为GD/1/96),根据测得的NS基因序列,设计一对特异性引物NS1U/NS1L,RT-PCR方法扩增该毒株的NS1基因。将酶切处理后的基因片段定向克隆到原核表达载体PET-32a( ),并进一步酶切分析和序列测定,鉴定出NS1基因的阳性重组子。阳性质粒转化E.coliBL 21(DE3)感受态细胞,在IPTG诱导下,经SDS-PAGE和Western-blotting分析鉴定,成功的在细菌包涵体中表达出45 kD的NS1融合蛋白。重组蛋白经Ni-NTA His.Bind Resins纯化。 |
| 关 键 词： | H5N1禽流感病毒 NS1蛋白 克隆 原核表达 纯化 |
| 文章编号： | 1007-8614（2007）03-0069-04 |
| 修稿时间： | 2007-03-15 |
Cloning of Nonstructural (NS1) Gene of H5N1 Avian Influenza Virus and Its Prokaryotic Expression in E.coli |
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| LIU Wei-long,JIAO Pei-rong,LIU Chang,Ahemaiti·Maimaiti,CHEN Hua-lan.Cloning of Nonstructural (NS1) Gene of H5N1 Avian Influenza Virus and Its Prokaryotic Expression in E.coli[J].Journal of Xinjiang Agricultural University,2007,30(3):69-72. | |
| Authors: | LIU Wei-long JIAO Pei-rong LIU Chang Ahemaiti·Maimaiti CHEN Hua-lan |
| Institution: | 1. College of Animal Medicine, Xinjiang Agricultural University, Urmuqi 830052; 2. Key Laboratory of Animal Influenza of Ministry of Agriculture, Harbin Veterinary Research Institute of CAAS,Harbin 150001 ;3. College of Animal Sciences and Medicine,Inner Mongolia Agricultural University, Huhehot,010018 |
| Abstract: | According to the sequence of nonstructural protein(NS1) gene of A/Goose/Guangdong/1/96,a pair of primers(NS1U/NS1L) was designed,NS1 gene was amplified by RT-PCR,was cloned directionaly into the PET-32a( ),the constructed recombinant plasmid was analyzed.The positive plasmids were tansformed into E.coli BL21(DE3),and induced with IPTG.Analysis of SDS-PAGE and Western-blotting showed that recombinant fusion protein was successfully expressed in inclusion,its moleculer weight was 45 kD.The fusion protein was purified by affinity chromatograph. |
| Keywords: | H5N1 avian influenza virus NS1 protein cloning Prokaryotic Expression purification |
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