Legerski R J, Brown D B, Peterson C A, Robberson D L
Proc Natl Acad Sci U S A. 1984 Sep;81(18):5676-9. doi: 10.1073/pnas.81.18.5676.
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.
已开发出一种检测方法，通过显微注射源自HeLa细胞的细胞质聚腺苷酸（poly(A)+）RNA，以非预定DNA合成来衡量，其中着色性干皮病细胞的切除修复缺陷可得到短暂互补。已对着色性干皮病的四个不同互补组进行了检测。A组和G组显示出互补，而D组和F组则没有。显微注射后每组细胞的存活率约为75%。根据接近正常的非预定DNA合成判断，A组和G组中约10%-25%的存活细胞得到了互补。将细胞质聚腺苷酸（poly(A)+）RNA在15%-30%的非变性蔗糖梯度上进行分级分离，随后对各个级分进行显微注射，结果表明，与着色性干皮病A组和G组互补的修复mRNA分别在约11S和12S处沉降。该检测方法在DNA修复基因的克隆和生化分析中应具有很大的实用性。
