| 含PRRS病毒ORF5的伪狂犬病病毒TK基因缺失转移载体的构建 |
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| 引用本文： | 仇华吉,周彦君.含PRRS病毒ORF5的伪狂犬病病毒TK基因缺失转移载体的构建[J].动物医学进展,2000,21(4):46-49. |
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| 作者姓名： | 仇华吉 周彦君 |
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| 作者单位： | 哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨 |
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| 摘 要： | 提取伪狂犬病病毒（ＰＲＶ）ＢａｒｔｈａＫ６１株基因组ＤＮＡ，用限制性内切酶ＫｐｎＩ充分消化，回收５．９ｋｂ片段（Ｊ片段），将其克隆于质粒ｐＵＣ１１９ ＫｐｎＩ位点上，获得ｐＢＫＪ。用两对针对ＰＲＶ ＴＫ基因的特异性引物对重组质粒进行ＰＣＲ鉴定，证明其中含有ＰＲＶ ＴＫ基因。然后用ＫｐｎＩ、ＰｓｔＩ和ＢａｍＨＩ等限制性内切酶对其进行酶切分析，确定了克隆片段的物理图谱。进一步研究证实ＴＫ基因位于其中的
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| 关 键 词： | 伪狂犬病病毒 TK基因 基因缺失 转移载体 猪 |
Construction of the Thymidine Kinase Gene Deleted Pseudorabies Virus Transfer Vector Containing the ORF5 of Porcine Reproductive and Respiratory Syndrome Virus |
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| QIU Hua ji,ZHOU Yan jun,KONG Ling da,ZHANG Shao jie,QIAN Ping,WANG Liu,TONG Guang zhi.Construction of the Thymidine Kinase Gene Deleted Pseudorabies Virus Transfer Vector Containing the ORF5 of Porcine Reproductive and Respiratory Syndrome Virus[J].Progress In Veterinary Medicine,2000,21(4):46-49. |
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| Authors: | QIU Hua ji ZHOU Yan jun KONG Ling da ZHANG Shao jie QIAN Ping WANG Liu TONG Guang zhi |
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| Institution: | QIU Hua ji,ZHOU Yan jun,KONG Ling da,ZHANG Shao jie,QIAN Ping,WANG Liu,TONG Guang zhi * |
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| Abstract: | Pseudorabies virus (PRV) genome DNA was extracted and digested by restriction endonuclease KpnI. The KpnI fragment J of 5.9 kb was shown to contain thymidine kinase (TK) gene by polymerase chain reaction (PCR), and it was cloned to pUC119, resulting a recombinant pBTK5.9. The restriction and PCR analyses confirmed the TK gene was located in its PstI KpnI 2.6 kb fragment. The minor fragment was then subcloned to produce recombinant pBTK2.6. Porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 under the control of CMV promotor was inserted into the pBTK2.6. The resulting transfer plasmid pBdTK CMV ORF5 will be used for devolping TK - and gE - deletd recombinant PRV expressing PRRSV ORF5. |
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| Keywords: | pseudorabies virus porcine reproductive and respiratory syndrome virus thymidine kinase gene transfer vector8 |
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