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| 副猪嗜血杆菌Plp4蛋白的原核表达及免疫原性分析 | |
| 引用本文： | 朱晓凯,赵福广,李建军,胡明明,李艳玲,张艳禾,谢芳,王春来.副猪嗜血杆菌Plp4蛋白的原核表达及免疫原性分析[J].中国预防兽医学报,2012,34(5):415-417. |
| 作者姓名： | 朱晓凯 赵福广 李建军 胡明明 李艳玲 张艳禾 谢芳 王春来 |
| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/细菌病研究室,黑龙江哈尔滨150001;吉林农业大学生命科学学院,吉林长春130118 2. 吉林农业大学生命科学学院,吉林长春,130118 3. 中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室/细菌病研究室,黑龙江哈尔滨,150001 |
| 基金项目： | 兽医生物技术国家重点实验室自主研究课题副猪嗜血杆菌病灭活苗的研制(SKLVBP201015) |
| 摘 要： | 为原核表达副猪嗜血杆菌P1p4蛋白,本研究通过PCR方法扩增P1p4全长基因并克隆于pET-28a(+)载体中,将重组质粒转化BL21(DE3)感受态中,采用0.4 mM IPTG经22℃诱导表达了35 ku的重组蛋白.经western blot试验证明Plp4蛋白具有良好的反应原性,免疫6周龄昆明小鼠制备免疫血清,ELISA检测表明制备的抗血清效价在1∶15000以上,表明P1p4蛋白具有良好的免疫原性. |
| 关 键 词： | 副猪嗜血杆菌 表达 Plp4基因 免疫原性 |
Prokaryotic expression of Plp4 gene from Haemophilus parasuis and antigenic analysis |
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| ZHU Xiao-kai , ZHAO Fu-guang , LI Jian-jun , HU Ming-ming , LI Yan-ling , ZHANG Yan-he , XIE Fang , WANG Chun-lai.Prokaryotic expression of Plp4 gene from Haemophilus parasuis and antigenic analysis[J].Chinese Journal of Preventive Veterinary Medicine,2012,34(5):415-417. | |
| Authors: | ZHU Xiao-kai ZHAO Fu-guang LI Jian-jun HU Ming-ming LI Yan-ling ZHANG Yan-he XIE Fang WANG Chun-lai |
| Institution: | 1(1.Division of Animal Bacteriosis,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Harbin 150001,China;2.College of Life Science,Jilin Agricultural University,Changchun 130118,China) |
| Abstract: | The full-length Plp4 gene was amplified from Haemphilus parasuis serotype 5 by PCR and cloned into vector pET-28a(+).The resultant recombinant plasmid was transformed into competent E.coli BL21(DE3) and the soluble recombinant plp4(rPlp4) which had a MW of 35 ku was expressed with IPTG induction at 22 ℃.The rPlp4 protein was recognized with anti-H.parasuis serum identified by western blot.The anti-Plp4 serum prepared in Kunming mice immunized by purified rPlp4 with a titer of 1∶15 000 revealed that the rPlp4 had high immunogenicity. |
| Keywords: | Haemophilus parasuis expression Plp4 gene immunogenicity |
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