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| 链球菌GapC基因重组痘苗病毒的构建及筛选与鉴定 | |
| 引用本文： | 李国华,李 贞,乔 军,陈创夫,才学鹏,孟庆玲,丁 波.链球菌GapC基因重组痘苗病毒的构建及筛选与鉴定[J].西北农业学报,2012,21(5):41-45. |
| 作者姓名： | 李国华 李 贞 乔 军 陈创夫 才学鹏 孟庆玲 丁 波 |
| 作者单位： | 1. 石河子大学,新疆地方与民族高发病教育部重点实验室,新疆石河子832003 2. 中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,兰州730046 3. 石河子大学,新疆地方与民族高发病教育部重点实验室,新疆石河子832003;中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,兰州730046 |
| 基金项目： | 兵团博士资金项目，石河子大学高层次人才专项项目 |
| 摘 要： | 为了研制链球菌性奶牛乳房炎的有效疫苗,首先用PCR方法扩增出停乳链球菌的GapC基因。其次将GapC基因连接到pSC11穿梭质粒上,构建pSC11-GapC重组穿梭质粒。然后在脂质体的介导下将pSC11-GapC转染已感染痘苗病毒的BHK-21细胞,使之与痘苗病毒基因在BHK-21细胞内进行同源重组,得到GapC重组痘苗病毒,用rVV-GapC表示。最后用蓝斑法连续筛选5次rVV-GapC重组毒,再用PCR和Western blot分别检测rVV-GapC的重组和表达情况。结果显示,PCR电泳检测到目的条带,测序结果与GenBank公布的标准序列同源性99%,Western blot试验证明GapC蛋白在重组毒中进行了表达。rVV-GapC构建成功。重组痘苗病毒的构建为链球菌性奶牛乳房炎疫苗的研究奠定基础。 |
| 关 键 词： | GapC基因 pSC11质粒 pSC11-GapC 痘苗病毒 rVV-GapC |
Construction, Screening and Identification of Recombinant Vaccinia Virus Expressing GapC Gene of Streptococcus |
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| LI Guohu,LI Zhen,QIAO Jun,CHEN Chuangfu,CAI Xuepeng,MENG Qingling and DING Bo.Construction, Screening and Identification of Recombinant Vaccinia Virus Expressing GapC Gene of Streptococcus[J].Acta Agriculturae Boreali-occidentalis Sinica,2012,21(5):41-45. | |
| Authors: | LI Guohu LI Zhen QIAO Jun CHEN Chuangfu CAI Xuepeng MENG Qingling and DING Bo |
| Institution: | 1(1.Key Laboratory of the High Incidence of Local and National Disease of Ministry of Education,Shihezi University, Shihezi Xinjiang 832003,China;2.Key Laboratory of National Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China) |
| Abstract: | In order to acquire the vaccine against the mastitis caused by Streptococcus, the GapC gene was firstly amplified from Streptococcus dysgalactiae. Secondly the gene was cloned into plasmid pSC11 to construct the recombinant plasmid pSC11 GapC. Thirdly, with the help of lipidosome, plasmid pSC11 GapC was transfected into the BHK 21cells, which had been infected by vaccinia virus. The aim is to make the virus and pSC11 GapC recombined randomly and get the recombinant, which is defined as rVV GapC. Lastly, rVV GapC was screened by blue plaque for 5 continuous generations, and then detected the GapC recombination and translation. The result showed that GapC was amplified by PCR, and sequence homology was 99% compared with the published sequence, and GapC protein was detected by western blot. So rVV GapC has been successfully constructed. The construction of rVV GapC laid the foundations for the research of Streptococcus vaccines. |
| Keywords: | GapC gene Plasmid pSC11 pSC11 GapC Vaccinia virus rVV GapC |
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