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| 表达禽流感HA基因的重组马立克氏病病毒的构建 | |
| 引用本文： | 周雪媚,霍惠玲,佘锐萍,李永清,冯国民,章振华,王宏俊.表达禽流感HA基因的重组马立克氏病病毒的构建[J].华北农学报,2007,22(4):168-171. |
| 作者姓名： | 周雪媚 霍惠玲 佘锐萍 李永清 冯国民 章振华 王宏俊 |
| 作者单位： | 1. 中国农业大学,动物医学院,北京,100094;河北省兽药监察所,河北,石家庄,050051 2. 河北省兽药监察所,河北,石家庄,050051 3. 中国农业大学,动物医学院,北京,100094 4. 北京市农林科学院畜牧兽医研究所,北京,100089 5. 江西农业大学,动物医学院,江西,南昌,330045 |
| 基金项目： | 河北省自然科学基金(C2005000654) |
| 摘 要： | 本试验首先用RT-PCR方法扩增出H9N2亚型禽流感病的HA基因,大小约1 700 bp,将其克隆到真核表达载体pcDNA中,并将该真核表达载体上的CMV启动子控制的含LacZ和HA基因表达盒克隆入马立克氏病病毒US2基因中,成功的构建了马立克氏病病毒的转移载体。然后,通过脂质体方法转染已感染马立克氏病病毒的鸡胚成纤维细胞(CEF)中,通过蓝斑筛选获得重组病毒。该重组病毒通过PCR方法鉴定证实其基因组中含有完整的HA基因。免疫酶反应和Western-blot证实重组病毒表达了禽流感HA蛋白,表达的HA蛋白具有血凝活性。 |
| 关 键 词： | 禽流感病毒 HA基因 重组马立克氏病病毒 |
| 文章编号： | 1000-7091（2007）04-0168-04 |
| 修稿时间： | 2007-04-12 |
Construction of Recombinant Marek's Disease Virus Expressing Avian Influenza Virus HA Gene |
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| ZHOU Xue-mei,HUO Hui-ling,SHE Rui-ping,LI Yong-qing,FENG Guo-min,ZHANG Zhen-hua,WANG Hong-jun.Construction of Recombinant Marek''''s Disease Virus Expressing Avian Influenza Virus HA Gene[J].Acta Agriculturae Boreali-Sinica,2007,22(4):168-171. | |
| Authors: | ZHOU Xue-mei HUO Hui-ling SHE Rui-ping LI Yong-qing FENG Guo-min ZHANG Zhen-hua WANG Hong-jun |
| Institution: | 1. Department of Veterinary Medicine, China Agricultural University, Beijing 100094, China ;2. Hebei Province Institute of Veterinary Drug Control, Shijiazhuang 050051, China ;3. Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100089, China; 4. Department of Veterinary Medicine,Jiangxi Agricultural University, Nanchang 330045, China |
| Abstract: | In this study,a 1.7 kb DNA fragment encoding the HA gene of Avian influenza virus H9N2 was obtained by RT-PCR and the sequence was cloned into the vector of pcDNA;Then the expression cassette including the HA gene of AIV and LacZ gene of Ecoli controlled by CMV promoter,were inserted into US2 gene of MDV to give a transfer vector.The complex of pUS2 HA and DOTAP was transfected into CEF infected MDV.The recombinant MDV were selected and purified by blue plague.The complete HA gene inserted into MDV was detected by PCR.Western-blot and immunostaining result demonstrated that HA protein of AIV was expressed by recombinant MDV. |
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