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人类腺嘌呤磷酸核糖转移酶在16号染色体上的基因座。

The locus for human adenine phosphoribosyltransferase on chromosome no. 16.

作者信息

Kahan B, Held K R, DeMars R

出版信息

Genetics. 1974 Dec;78(4):1143-56. doi: 10.1093/genetics/78.4.1143.

Abstract

Evidence for assigning the locus determining the structure of adenine phosphoribosyltransferase (APRT) to human chromosome No. 16 is presented. Hybrids of APRT-deficient mouse cells and of human fibroblasts having normal APRT were isolated by fusing the parental cells with Sendai virus, blocking de novo purine nucleotide synthesis with azaserine and selecting for hybrids that could use exogenous adenine. The hybrid clones that were studied had only APRT activity that was indistinguishable from human APRT with regard to electrophoretic migration and reaction with antibodies against the partially purified human enzyme. No. 16 was the only human chromosome consistently present in all of the clones, and in one clone, it was the only human chromosome detected. Selection against hybrid cells with 2,6-diaminopurine (DAP) yielded DAP-resistant survivors that lacked chromosome No. 16. One hybrid that originally had an intact No. 16 yielded adenine-utilizing subclones that lacked No. 16 but had a new submetacentric chromosome. The distribution of centromere-associated heterochromatin and the fluorescence pattern indicated that this chromosome consisted of a mouse telocentric chromosome and the long arm of No. 16. Cells having the submetacentric chromosome had human APRT. Both the enzyme and the chromosome were absent in DAP-resistant derivatives. These results suggest that the structure of APRT is defined by a locus on the long arm of human chromosome No. 16.

摘要

本文提供了将决定腺嘌呤磷酸核糖转移酶(APRT)结构的基因座定位于人类第16号染色体的证据。通过将亲代细胞与仙台病毒融合,用重氮丝氨酸阻断嘌呤核苷酸的从头合成,并选择能够利用外源性腺嘌呤的杂种细胞,分离出了APRT缺陷型小鼠细胞与具有正常APRT的人类成纤维细胞的杂种细胞。所研究的杂种克隆仅具有APRT活性,就电泳迁移率以及与针对部分纯化的人类酶的抗体的反应而言,该活性与人类APRT无法区分。16号染色体是所有克隆中始终存在的唯一人类染色体,并且在一个克隆中,它是唯一检测到的人类染色体。用2,6-二氨基嘌呤(DAP)对杂种细胞进行选择,产生了缺乏16号染色体的DAP抗性存活细胞。一个最初具有完整16号染色体的杂种产生了利用腺嘌呤的亚克隆,这些亚克隆缺乏16号染色体,但有一条新的亚中着丝粒染色体。着丝粒相关异染色质的分布和荧光模式表明,这条染色体由一条小鼠端着丝粒染色体和16号染色体的长臂组成。具有亚中着丝粒染色体的细胞具有人类APRT。在DAP抗性衍生物中,该酶和该染色体均不存在。这些结果表明,APRT的结构由人类第16号染色体长臂上的一个基因座决定。

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