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猪圆环病毒II型感染性DNA克隆的构建
  • ISSN号:1000-6850
  • 期刊名称:《中国农学通报》
  • 分类:S855.3[农业科学—临床兽医学;农业科学—兽医学;农业科学—畜牧兽医]
  • 作者机构:[1]北京农学院动物科学技术学院,北京102206
  • 相关基金:国家自然科学基金项目“微血管内皮细胞在猪圆环病毒II型感染中的作用”(30600442);北京市自然科学基金项目“猪圆环病毒II型感染对伪狂犬病疫苗免疫反应的影响”(6082008);北京市教委科技计划项目“嵌合猪圆环病毒疫苗株的构建”(KM200810020013).
中文摘要:

为了构建猪圆环病毒II型(PCV2)感染性DNA克隆,并为PCV2的突变体构建及其致病机理研究建立基础。用PCR方法从PCV2DNA质粒中扩增出PCV2全基因组片段,将其插入到pBluescript-SK载体中,构建出单拷贝PCV2DNA克隆,并进一步构建了双拷贝PCV2DNA克隆,将PCV2DNA克隆转染PK-15细胞,以获得PCV2拯救病毒,并初步测定了拯救病毒的体外增殖能力。结果显示:成功获得了PCV2拯救病毒,并能够在PK-15细胞中进行稳定传代,其TCID5。在连传5代后达到10^-6.05/mL,与其亲本病毒相近,显示出较高的感染滴度。研究结果表明,成功构建了具有较高感染性的PCV2感染性克隆,为PCV2的生物学特性及致病机理研究奠定了基础。

英文摘要:

The aim of this study was to construct an infectious DNA clone of Porcine Circovirus Type II (PCV2) and lay the foundations for the mutant construction and pathogenesis researches of the PCV2. The complete genome of PCV2 was amplified from the PCV2 DNA plasmid by PCR method, and inserted into the pBluescript-SK vector to construct the single copy of PCV2 DNA clone. Furthermore, the double copy of PCV2 DNA clone was developed. Then, the transfection of porcine kidney (PK)-15 cells with the PCV2 DNA clones was conducted to obtain the rescue viruses of PCV2, and the reproducibility of the rescue virus in vitro was determined. The results showed that the rescue viruses of PCV2 were acquired successfully and could propagate stably in PK-15 cells, its 50% tissue culture infectious doses (TCIDso) was 10-6~5/mL after 5 serial passages, which was close to its parent virus and demonstrated its relatively high infectious titer. The research results revealed that an infectious clone of PCV2 with relatively high infectivity was developed successfully, which established a foundation for further research on the virus biological characteristics and pathogenesis.

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期刊信息
  • 《中国农学通报》
  • 北大核心期刊(2008版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国农学会
  • 主编:石元春
  • 地址:北京市朝阳区麦子店街22号中国农学期刊处
  • 邮编:100025
  • 邮箱:edit@agri.org.cn
  • 电话:010-59194480
  • 国际标准刊号:ISSN:1000-6850
  • 国内统一刊号:ISSN:11-1984/S
  • 邮发代号:2-772
  • 获奖情况:
  • 全国农业核心期刊,中国科协优秀学术期刊,全国农业学会优秀期刊
  • 国内外数据库收录:
  • 中国中国科技核心期刊,中国北大核心期刊(2008版)
  • 被引量:94571