| 基于Flp重组酶介导的盒式交换HPRT位点定点转基因研究 |
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| 引用本文： | 郑敬民,贺艳,傅继梁.基于Flp重组酶介导的盒式交换HPRT位点定点转基因研究[J].分子细胞生物学报,2001,34(4):279-282. |
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| 作者姓名： | 郑敬民 贺艳 傅继梁 |
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| 作者单位： | 郑敬民（第二军医大学医学遗传学教研室,上海200433）
贺艳（第二军医大学医学遗传学教研室,上海200433）
傅继梁（第二军医大学医学遗传学教研室,上海200433） |
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| 基金项目： | 国家自然科学基金重点项目(39830363)资助. |
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| 摘 要： | 以电穿孔转染法将构建的交换质粒pF-HPRT-F3和Flp表达载体pCMV-Flp共转染已在基因组HPRT位点整合有交换盒F-Neo-F3结构的ES细胞F18,经HAT筛选得到HAT抗性ES细胞克隆;G418敏感性试验和基因组Southern杂交表明,所分析的12个HAT抗性克隆中有3个克隆发生了预期的盒式交换重组,转基因定点整合的相对效率为25%.这一结果表明,Flp重组酶是可以在HPRT位点有效地介导盒式交换反应的,我们提出的利用Flp重组酶介导的盒式交换反应建立基于HPRT位点的转基因定点整合体系的策略是可行的.
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| 关 键 词： | HPRT基因 重组酶介导的盒式交换 Flp 重组酶 |
| 修稿时间： | 2001年6月20日 |
THE STUDY OF A HPRT LOCUS-SPECIFIC TRANSGENIC METHOD BASED ON FLP RECOMBINASE MEDIATED CASSETTE EXCHANGE |
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| ZHENG Jing Min HE Yan.THE STUDY OF A HPRT LOCUS-SPECIFIC TRANSGENIC METHOD BASED ON FLP RECOMBINASE MEDIATED CASSETTE EXCHANGE[J].Journal of Molecular Cell Biology,2001,34(4):279-282. |
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| Authors: | ZHENG Jing Min HE Yan |
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| Abstract: | Exchange plasmid pF-HPRT-F3 and Flp expression plasmid pCMV-Flp were constructed and then introduced using electroporation system into F18 ES cell line, which have an exchange cassette F-Neo-F3 at HPRT locus. After HAT selection, HAT resistant clones were obtained. Then G418 sensitivity test and Southern blotting were
carried out to screen RMCE recombinants. The results indicated that RMCE had taken place in three of 12 HAT resistant clones. The frequency is 25%. The result demonstrates that it is realizable to introduce transgene to HPRT locus by using Flp recombinase mediated cassette exchange reaction. |
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| Keywords: | HPRT gene recombinase mediated cassette exchange Flp recombinase |
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