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以人类X染色体的次黄嘌呤磷酸核糖基转移酶(HPRT)区域为模型对染色体介导的基因转移进行研究。

Investigation of chromosome-mediated gene transfer using the HPRT region of the human X chromosome as a model.

作者信息

Pritchard C A, Goodfellow P N

机构信息

Laboratory of Human Molecular Genetics, Imperial Cancer Research Fund, London, UK.

出版信息

Genes Dev. 1987 Apr;1(2):172-8. doi: 10.1101/gad.1.2.172.

Abstract

A panel of over 50 hybrid cells containing varying portions of the long arm of the human X chromosome have been obtained by chromosome-mediated gene transfer (CMGT) of human chromosomes to mouse cells deficient in HPRT. This panel is used to investigate the size and integrity of transfected human chromosome fragments and also to examine the effect of including a selectable DNA plasmid in the transfection mix. Chromosomal rearrangements are found to be generated in the chromosome transfer process, and the human X centromeric region is detected in the transfected cells at an unusually high frequency. Extensive lengths of X chromosome DNA are transferred intact, suggesting potential uses of CMGT in cloning large genes and loci for which only the chromosomal map position is known.

摘要

通过将人类染色体进行染色体介导的基因转移(CMGT)到缺乏次黄嘌呤磷酸核糖转移酶(HPRT)的小鼠细胞中,已获得了一组包含人类X染色体长臂不同部分的50多个杂交细胞。该细胞组用于研究转染的人类染色体片段的大小和完整性,同时也用于检验在转染混合物中加入一个可选择的DNA质粒的效果。发现在染色体转移过程中会产生染色体重排,并且在转染细胞中以异常高的频率检测到人类X染色体着丝粒区域。大量的X染色体DNA完整地被转移,这表明CMGT在克隆仅知道染色体图谱位置的大基因和基因座方面具有潜在用途。

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