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通过显微注射酵母人工染色体（YAC）DNA将人类次黄嘌呤磷酸核糖转移酶（HPRT）基因和甘氨酰胺核糖核苷酸转甲酰基酶（GART）基因从酵母转移至哺乳动物细胞。

Transfer of the human HPRT and GART genes from yeast to mammalian cells by microinjection of YAC DNA.

作者信息

Gnirke A, Huxley C

机构信息

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

出版信息

Somat Cell Mol Genet. 1991 Nov;17(6):573-80. doi: 10.1007/BF01233622.

DOI:10.1007/BF01233622
PMID:1767336
Abstract

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.

摘要

将包含可选择人类基因的两条酵母人工染色体（YAC）的DNA通过显微注射转移到组织培养中的啮齿动物细胞中。如其他地方所述，跨越45 kb的人类次黄嘌呤磷酸核糖基转移酶（HPRT）基因包含在660 kb的YAC yHPRT上。如先前所述，跨越约40 kb的人类磷酸核糖甘氨酰胺甲酰基转移酶（GART）基因包含在590 kb的YAC yGART2上。从脉冲场凝胶中分离出YAC DNA，并显微注射到可以选择人类HPRT和GART基因的哺乳动物细胞中。选择的细胞系包含完整的人类基因。一些细胞系包含在同一染色体位置整合的多个基因拷贝。YAC yGART2无法从大小相似的天然酵母染色体中纯化出来，导入人类GART基因的细胞系除了人类DNA外还包含可变数量的酵母DNA。

相似文献

1
Transfer of the human HPRT and GART genes from yeast to mammalian cells by microinjection of YAC DNA.通过显微注射酵母人工染色体（YAC）DNA将人类次黄嘌呤磷酸核糖转移酶（HPRT）基因和甘氨酰胺核糖核苷酸转甲酰基酶（GART）基因从酵母转移至哺乳动物细胞。
Somat Cell Mol Genet. 1991 Nov;17(6):573-80. doi: 10.1007/BF01233622.
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Nucleic Acids Res. 2000 Sep 1;28(17):E81. doi: 10.1093/nar/28.17.e81.
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Construction and validation of yeast artificial chromosome contig maps by RecA-assisted restriction endonuclease cleavage.通过RecA辅助的限制性内切酶切割构建和验证酵母人工染色体重叠群图谱。
Proc Natl Acad Sci U S A. 1998 Sep 15;95(19):11318-23. doi: 10.1073/pnas.95.19.11318.
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Delivery of bacterial artificial chromosomes into mammalian cells with psoralen-inactivated adenovirus carrier.
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Nucleic Acids Res. 1997 May 15;25(10):1950-6. doi: 10.1093/nar/25.10.1950.
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Mol Cell Biol. 1993 Sep;13(9):5469-78. doi: 10.1128/mcb.13.9.5469-5478.1993.
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