| 通用伪狂犬病病毒转移载体的构建 |
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| 引用本文： | 仇华吉,童光志,周彦君,李昌文,钱平,孔令达.通用伪狂犬病病毒转移载体的构建[J].中国预防兽医学报,2000(Z1). |
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| 作者姓名： | 仇华吉 童光志 周彦君 李昌文 钱平 孔令达 |
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| 摘 要： | 克隆伪狂犬病病毒（ＰＲＶ） Ｂａｒｔｈａ－Ｋ６１株基因组的ＫｐｎⅠ Ｊ片段，然后亚克隆其中的ＫｐｎⅠ－ ＰｓｔⅠ片段，缺失重组质粒中的ＥｃｏＲⅠ位点和ＮｏｔⅠ－Ｈｉｎｄ Ⅲ片段；再用ＡｃｃⅠ切去３７８ｂｐ，在此缺失位置插入来源于ｐＣＲ３－Ｕｎｉ的ＣＭＶ启动子、多克隆位点和ＢＧＨ ｐｏｌｙＡ信号，构建了通用 ＰＲＶ转移载体 ｐＢｄＴＫ－Ｕｎｉ。此转移载体为改造 Ｂａｒｔｈａ－Ｋ６１株及开发二价或多价基因工程疫苗提供了有力工具。
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| 关 键 词： | 伪狂犬病病毒 TK基因 转移载体 |
Construction of a Universal Pseudorabies Virus Transfer Vector |
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| QIU Huaji,ZHOU Yanjun,LI Changwen,QIAN Ping,KONG Lingda,TONG Guangzhi.Construction of a Universal Pseudorabies Virus Transfer Vector[J].Chinese Journal of Preventive Veterinary Medicine,2000(Z1). |
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| Authors: | QIU Huaji ZHOU Yanjun LI Changwen QIAN Ping KONG Lingda TONG Guangzhi |
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| Abstract: | Pseudorabies virus(PRV)genome DNA was extracted and digested by the restriction endonuclease(RE) KpnI.The KpnI frag- ment J of 5 .9 kb was shown to contain thymidine kinase (TK) gene by polymerase chain reaction(PCR) , and it was cloned into pUC119, resulting in a recombinant pBTK5. 9. The RE and PCR analyses confirmed that the TK gene was located in the 2. 6 kb PstI- Kpn I fragment of the former fragment. This minor fragment was then subcloned to produce recombinant pBTK2. 6. A fragment containing the immediate early promoter of cytomegalovirus and bovine growth hormone polyadenylation signal derived from pCR3-Uni, a eukaryotic expression plasmid, was amplified and inserted into the AccI-deleted locus of pBTK2. 6. The resulting transfer vector pBdTK-Uni will be useful for developing TK-and gE-deleted recombinant PRV to express foreign gene(s). |
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| Keywords: | Pseudorabies virus Thymidine kinase gene Transfer vector |
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