TRANSFER OF THE HUMAN HPRT AND GART GENES FROM YEAST TO MAMMALIAN-CELLS BY MICROINJECTION OF YAC DNA

被引：15

作者：

GNIRKE, A

HUXLEY, C

机构：

[1] Department of Genetics, Washington University School of Medicine, St. Louis, 63110, Missouri

来源：

SOMATIC CELL AND MOLECULAR GENETICS | 1991年 / 17卷 / 06期

关键词：

D O I：

10.1007/BF01233622

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：

DNA of two yeast artificial chromosomes (YACs) containing selectable human genes was transferred by microinjection to rodent cells in tissue culture. The human hypoxanthine phosphoribosyltransferase (HPRT) gene, spanning 45 kb, is contained on the 660-kb YAC yHPRT as described elsewhere. The human phosphoribosylglycinamide formyltransferase (GART) gene, spanning approximately 40 kb, is contained on the 590-kb YAC yGART2 as described previously. YAC DNA was isolated from pulsed-field gels and microinjected into mammalian cells in which the human HPRT and GART genes can be selected. The cell lines that were selected contain the entire human genes. Some of the cell lines contain multiple copies of the genes integrated at the same chromosomal position. The YAC yGART2 could not be purified away from natural yeast chromosomes of similar size, and the cell lines into which the human GART gene was introduced contain variable amounts of yeast DNA in addition to the human DNA.

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页码：573 / 580

页数：8

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