INVESTIGATION OF COELECTROPORATION AS A METHOD FOR INTRODUCING SMALL MUTATIONS INTO EMBRYONIC STEM-CELLS

被引：14

作者：

DAVIS, AC

WIMS, M

BRADLEY, A

机构：

[1] Institute for Molecular Genetics, Baylor College of Medicine, Texas Medical Center, Houston

来源：

MOLECULAR AND CELLULAR BIOLOGY | 1992年 / 12卷 / 06期

关键词：

D O I：

10.1128/MCB.12.6.2769

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have investigated coelectroporation as a method for introducing minor genetic changes into specific genes in embryonic stem cells. A selectable marker (neo) and a targeting replacement vector designed to insert a 4-bp insertion into exon 3 of the mouse hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene were coelectroporated into embryonic stem cells and selected in G418 and 6-thioguanine (6-TG). HPRT-negative clones were obtained at a frequency of approximately 1 per 520 G418r clones. Southern analysis and the polymerase chain reaction were used to demonstrate that 3 of 36 of the 6-TG-resistant clones had the desired 4-bp insertion without any other disruption of the HPRT locus. Initial studies indicated that the other 33 6-TG-resistant clones probably resulted from the targeted intergration of a concatemer containing both the targeting construct and the selectable neo gene.

引用

页码：2769 / 2776

页数：8

共 29 条

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