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猪捷申病毒Swine/CH/IMH/03全长cDNA克隆的构建 被引量：2

Construction of a full-length cDNA clone of PTV Swine/CH/IMH/03
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摘要 为构建猪捷申病毒(Porcine testhovirus,PTV)感染性克隆,进行病毒基因组结构、功能及新型疫苗研究,根据NCBI发表的PTV Swine/CH/IMH/03株及Talfan株核苷酸序列,设计并合成了4对特异性引物,PCR扩增并克隆覆盖PTV Swine/CH/IMH/03全长基因组的4个片段。经过反复拼接和测序,最终将PTV全长cDNA定向克隆到pBluescript SK(+)中。PCR、双酶切及测序鉴定证明PTV全长cDNA克隆构建成功。与Swine/CH/IMH/03亲本株及NCBI登录的其他PTV毒株序列比对发现,该克隆只有3个位点存在核苷酸及氨基酸突变,分别位于2A、2C及3D中。 According to the published nucleotide sequence of PTV Swine/CH/IMH/03 and Talfan, 4 pairs of primers were designed for RT-PCR to amplify 4 overlapping fi＇agments covering PTV swine/CH/IMH/03 genome. These fragments were subcloned to pMD18-T and pBluescript SK （＋） and assembled. A PTV full-length cDNA clone pSK-PTV was cloned.to pBluescript SK （＋）, and confirmed by PCR, restrictive digestion and sequencing. Comparing to the published seuqneces of most PTV strains, this clone possessed only 3 nucleotides and amino acids mutations located in 2A, 2C and 3D genes respectively. The successful construction of PTV full-length cDNA clone provided a useful platform for PTV reverse genetics and the offspring study.
作者 吴波平 冯力 时洪艳 陈建飞 孙东波 高秀春 白兴华
机构地区 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
出处 《中国预防兽医学报》 CAS CSCD 北大核心 2007年第12期929-933,942,共6页 Chinese Journal of Preventive Veterinary Medicine
关键词 PTV 全长CDNA克隆 感染性克隆 PTV full-length cDNA infectious clone
分类号 S852.659.6 [农业科学—基础兽医学]
  • 相关文献

参考文献12

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共引文献1

同被引文献17

  • 1吴波平,冯力,时洪艳,陈建飞,孙东波,高秀春,白兴华.猪捷申病毒3D蛋白的原核高效表达及其反应活性[J].中国兽医科学,2007,37(2):131-134. 被引量：7
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引证文献2

二级引证文献10

中国预防兽医学报
2007年 第12期

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