Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA.
Transient complementation of xeroderma pigmentosum cells by microinjection of poly(A)+ RNA.
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通过显微注射聚 (A) RNA 瞬时补充着色性干皮病细胞。
DOI：
10.1073/pnas.81.18.5676
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发表时间：
1984
影响因子：
11.1
通讯作者：
D. Robberson
中科院分区：
文献类型：
--
作者：
R. Legerski;D. B. Brown;C. Peterson;D. Robberson
已经开发出一种检测方法，通过显微注射源自 HeLa 细胞的细胞质 Poly(A)+ RNA，可暂时补充着色性干皮病细胞的切除修复缺陷，这是通过非计划性 DNA 合成来测量的。已经测定了着色性干皮病的四个不同的互补组。 A 组和 G 组显示出互补性，而 D 组和 F 组则没有。显微注射后各组细胞的存活率大约等于 75%。根据接近正常的计划外 DNA 合成判断，A 组和 G 组中大约 10-25% 的存活细胞得到了补充。在 15-30% 非变性蔗糖梯度上对细胞质 Poly(A)+ RNA 进行分级分离，并随后对各个级分进行显微注射，表明补充着色性干皮病 A 组和 G 组的修复 mRNA 分别沉积在大约 11 S 和 12 S 处。该测定在 DNA 修复基因的克隆和生化分析中应该具有很大的用途。
An assay has been developed in which excision repair deficiency of xeroderma pigmentosum cells is transiently complemented, as measured by unscheduled DNA synthesis, by microinjection of cytoplasmic poly(A)+ RNA derived from HeLa cells. Four different complementation groups of xeroderma pigmentosum have been assayed. Groups A and G showed complementation, whereas groups D and F did not. Survival for cells in each of the groups subsequent to microinjection was approximately equal to 75%. Approximately 10-25% of surviving cells from groups A and G were complemented, as judged by near-normal unscheduled DNA synthesis. Fractionation of cytoplasmic poly(A)+ RNA on a 15-30% nondenaturing sucrose gradient and subsequent microinjection of the individual fractions indicate that repair mRNAs that complement xeroderma pigmentosum groups A and G sediment at approximately 11 S and 12 S, respectively. This assay should be of great utility in the cloning and biochemical analysis of DNA repair genes.