| 猪细小病毒BQ-C毒株VP2基因的鉴定及表达 |
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| 引用本文： | 刘朝霞,刘永刚,毕可东,周顺,崔尚金.猪细小病毒BQ-C毒株VP2基因的鉴定及表达[J].中国畜牧兽医,2013,40(11):50-53. |
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| 作者姓名： | 刘朝霞 刘永刚 毕可东 周顺 崔尚金 |
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| 作者单位： | 1. 青岛农业大学动物科技学院, 山东青岛 266109;2. 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, 猪传染病研究室, 黑龙江哈尔滨 150001 |
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| 基金项目： | 国家自然科学基金(31001069、31172349、31172341)。 |
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| 摘 要： | 本研究从临床发病猪采集病料,以PK15传代细胞进行病毒分离,通过PCR、血液凝集试验(HA)和电镜鉴定为1株猪细小病毒(porcine parvovirus,PPV),命名为BQ-C。对分离的毒株进行测序,结果显示该毒株与GenBank登录的PPV BQ株同源性为100%。为获得该毒株结构蛋白VP2基因的表达产物,将VP2基因片段插入到原核表达载体pET-30a(+),得到表达重组质粒。经双酶切和测序鉴定,将重组质粒转化大肠杆菌BL21(DE3)中进行表达。SDS-PAGE结果表明,获得的重组蛋白分子质量约为71.5 ku,与预期大小相符。Western blotting结果显示获得的重组蛋白能与PPV阳性血清特异性结合,表明重组蛋白具有良好的反应原性。结果表明,本研究成功分离了1株PPV BQ-C株,且表达的VP2重组蛋白可用于PPV血清学诊断和疫苗的研发。
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| 关 键 词： | 猪细小病毒 重组VP2蛋白 原核表达 反应原性 |
| 收稿时间： | 2013-04-10 |
Identification and Expression of VP2 Gene of Porcine Parvovirus BQ-C Isolate |
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| LIU Zhao-xia,LIU Yong-gang,BI Ke-dong,ZHOU Shun,CUI Shang-jin.Identification and Expression of VP2 Gene of Porcine Parvovirus BQ-C Isolate[J].China Animal Husbandry & Veterinary Medicine,2013,40(11):50-53. |
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| Authors: | LIU Zhao-xia LIU Yong-gang BI Ke-dong ZHOU Shun CUI Shang-jin |
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| Institution: | 1. College of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China;2. Division of Swine Infectious Disease, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China |
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| Abstract: | The BQ-C isolate of porcine parvovirus (PPV) was propagated on PK15 cell, and identified by polymerase chain reaction (PCR), haemagglutination test (HA) and electron microscope. To express the VP2 structural protein of PPV BQ-C strain, the VP2 gene was cloned into pET-30a(+) and expressed in E.coli BL21(DE3). SDS-PAGE analysis showed that the recombinant protein was about 71.5 ku. Western blotting assay confirmed that the protein was able to specifically react with PPV positive serum. In this study, the BQ-C isolate of PPV was successfully obtained, and the recombinant VP2 protein could be used for the diagnosis of PPV and vaccine development. |
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| Keywords: | porcine parvovirus recombinant VP2 protein prokaryotic expression immunoreactivity |
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