| 含E.coli F因子的马立克氏病病毒1型（MDV1）转移载体的构建及初步应用 |
| |
| 引用本文： | 邱亚峰,葛菲菲,陈溥言.含E.coli F因子的马立克氏病病毒1型（MDV1）转移载体的构建及初步应用[J].中国农业科学,2006,39(11):2341-2346. |
| |
| 作者姓名： | 邱亚峰 葛菲菲 陈溥言 |
| |
| 作者单位： | 南京农业大学动物医学院动物疫病诊断与免疫重点实验室,南京,210095 |
| |
| 基金项目： | 国家高技术研究发展计划(863计划) |
| |
| 摘 要： | 【目的】为了构建MDV1细菌人工染色体，以便快速有效地进行重组病毒的筛选，进行了含E.coli F因子的马立克氏病病毒1型（MDV1）转移载体的构建。【方法】利用DNA重组技术，构建了含有E.coli F因子的马立克氏病病毒1型（MDV1）转移载体，利用脂质体，将线性化的转移载体转染入已感染CVI988病毒的CEF，用MX-HAT药物筛选3代以后，用X-gal染色,挑取阳性克隆。【结果】经测序分析发现，人工体外合成的单一loxP位点，序列正确，另外，同源重组左臂和右臂序列正确，并且相对连接。利用SphⅠ和NheⅠ双酶切，鉴定出含有同向LoxP位点的转移载体，将其命名为pUS-BGS。将该载体用NheⅠ线性化后，用脂质体转染已感染CVI988病毒的CEF，用MX-HAT药物筛选3代以后，X-gal染色，可以观察到呈现蓝色的重组病毒。【结论】本研究报道了含有E.coli F因子以及两同向的loxP位点的MDV1转移载体的构建，利用该载体可以有效地进行重组病毒的筛选，为下一步MDV1细菌人工染色体的构建奠定了基础。
|
| 关 键 词： | 马立克氏病病毒 细菌人工染色体 F因子 |
| 收稿时间： | 2005-7-3 |
| 修稿时间： | 2005年7月3日 |
Construction of Transfer Vector of Marek's Disease Virus Serotype 1 (MDV1) Containing F Factor of Eschericha coli and its Primary Application |
| |
| QIU Ya-feng,GE Fei-fei,CHEN Pu-yan.Construction of Transfer Vector of Marek''''s Disease Virus Serotype 1 (MDV1) Containing F Factor of Eschericha coli and its Primary Application[J].Scientia Agricultura Sinica,2006,39(11):2341-2346. |
| |
| Authors: | QIU Ya-feng GE Fei-fei CHEN Pu-yan |
| |
| Abstract: | AIM] To construct the bacterial artificial chomosome containing the MDV1 genome, and further product fast and efficiently the recombinant virus of MDV1 by using BAC vector.MAIN METHORD] Using the DNA recombinant techniques, the transferred vector containing E.coli F factor was constructed. The linearized vector was transfected into CEF which was infected by CVI988 virus. After three-cycle selection by MX-HAT, the infected CEF was stained with X-gal. MAIN RESULT] The sequence analysis shows that the LoxP site synthesized in vitro was right,and the sequences of the homologous recombinant left arm and right arm are right. The two arms were ligated with the reversed oritation.The positive plasmid ,in which two LoxP sites had the same orientation, was identified by SphⅠand NheⅠ,named pUS-BGS. This linearized vector by NheⅠ was transfected into CEF which was infected by CVI988 virus. After three-cycle selection by MX-HAT, the infected CEF was stained with X-gal，blue plagues were appeared. MAIN CONCLUSION] This research first reports that the transferred vector containing the E.coli F factor and two LoxP sites which had the same orientation, was constructed. The result shows that this vector is efficient for the selection of recombinant MDV1. So this gives the basis for construction of the bacterial artificial chromosome of MDV1. |
| |
| Keywords: | Marek’s disease virus bacterial artificial chromosome F factor |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《中国农业科学》浏览原始摘要信息 |
| 点击此处可从《中国农业科学》下载免费的PDF全文 |
