| 马动脉炎病毒GL、M和N蛋白主要抗原域的原核表达及抗原性鉴定 |
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| 引用本文： | 王贵宾,郭巍,赵立平,李红梅,赵敏,相文华.马动脉炎病毒GL、M和N蛋白主要抗原域的原核表达及抗原性鉴定[J].兽医大学学报,2012(7):958-961. |
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| 作者姓名： | 王贵宾 郭巍 赵立平 李红梅 赵敏 相文华 |
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| 作者单位： | [1]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001 [2]东北林业大学生命科学学院,黑龙江哈尔滨150040 |
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| 基金项目： | 中央级公益性科研院所基本科研业务费专项（ZGK201007） |
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| 摘 要： | 采用RT—PCR方法从EAVBucyrus株扩增了截短的GL、M和全长的N基因片段，将三者分别克隆到表达载体pGEX-6p-1中，测序验证后转化Rosetta（DE3）宿主菌中经IPTG诱导表达，诱导后菌体裂解物经SDS-PAGE分析。试验结果表明，重组菌表达出约35000、34000和38000的特异性条带，通过条件优化及切胶纯化获得较高浓度的目的蛋白。经Western-blot和间接ELISA分析，纯化的蛋白只与抗马动脉炎病毒阳性血清发生特异性反应，证实该蛋白具有良好的反应原性和特异性。
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| 关 键 词： | 马动脉炎病毒 原核表达 抗原性 |
Prokaryotic expression of the major antigenic domain of equine arteritis GL, M and N protein and antigenicity of their expressed product |
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| Institution: | WANG Gui-bin , GUO Wei , ZHAO Li-ping , LI Hong-mei , ZHAO Min , XINAG Wen-Hua (1. Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China ; 2. College of Life Science, Northeast Forestry University, Harbin 150040, China) |
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| Abstract: | The gene of truncated GL and M and the nucleocapsid protein gene were amplified from equine arteritis vi- rus（EAV） Bucyrus strain by RT-PCR and coloned into the pGEX-6p-1 expression vector. After sequencing,the re- combinant plasmid were transformed into Rosetta（DE3）. The transformed bacteria were induced by IPTG and pro- duced a recombinant protein of 35 000,34 000 and 38 000 after purified. The purified protein interacted well with the positive serum against EAV in a Western-blot and indirect ELISA. There results showed that the recombinant pro- tein had good antigenicity and speificity. |
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| Keywords: | EAV prokaryotic expression antigenicity |
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