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腺苷激酶作为体细胞遗传学中的一种新型选择性标记:腺苷激酶缺陷型小鼠细胞系和含腺苷激酶的人-鼠杂交细胞系的分离

Adenosine kinase as a new selective marker in somatic cell genetics: isolation of adenosine kinase--deficient mouse cell lines and human--mouse hybrid cell lines containing adenosine kinase.

作者信息

Chan T S, Creagan R P, Reardon M P

出版信息

Somatic Cell Genet. 1978 Jan;4(1):1-12. doi: 10.1007/BF01546489.

Abstract

A new selective system for isolating somatic cell hybrids, using adenosine kinase as the selective marker, has been developed. The selective medium for forward selection (to select for cells containing adenosine kinase) contains alanosine, adenosine and uridine. To survive in the presence of alanosine, cells must have adenosine kinase in order to utilize exogenous adenosine as the sole source of AMP. Uridine is added to the selective medium to prevent the toxic effects of adenosine on cultured mammalian cells. The selective medium for reverse selection (to select for cells lacking adenosine kinase) contains 2-fluoroadenosine, an analogue of adenosine, which is converted to a toxic nucleotide by the action of adenosine kinase. Mouse mutant cell lines deficient in adenosine kinase have been derived. Human--mouse hybrid cells containing the kinase have been prepared from one of these mutant lines. Karyotype data of these hygrid lines and their adenosine kinase-minus sublines are consistent with assignment by others of the human gene for adenosine kinase on chromosome 10.

摘要

已开发出一种新的用于分离体细胞杂种的选择系统,该系统使用腺苷激酶作为选择标记。正向选择(用于选择含有腺苷激酶的细胞)的选择培养基含有丙氨酸、腺苷和尿苷。为了在丙氨酸存在下存活,细胞必须具有腺苷激酶,以便利用外源性腺苷作为AMP的唯一来源。向选择培养基中添加尿苷以防止腺苷对培养的哺乳动物细胞产生毒性作用。反向选择(用于选择缺乏腺苷激酶的细胞)的选择培养基含有2-氟腺苷,它是腺苷的类似物,在腺苷激酶的作用下会转化为有毒核苷酸。已获得缺乏腺苷激酶的小鼠突变细胞系。已从这些突变系之一制备出含有该激酶的人-鼠杂种细胞。这些杂种细胞系及其不含腺苷激酶的亚系的核型数据与其他人将人类腺苷激酶基因定位于10号染色体的结果一致。

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