从分离的uvrA⁺和uvrB⁺/uvrC⁺基因产物重建大肠杆菌修复内切核酸酶活性。
Reconstitution of an Escherichia coli repair endonuclease activity from the separated uvrA+ and uvrB+/uvrC+ gene products.
作者信息
Seeberg E
出版信息
Proc Natl Acad Sci U S A. 1978 Jun;75(6):2569-73. doi: 10.1073/pnas.75.6.2569.
Abstract
An in vitro complementation assay has been used for partial purification of uvrA+, uvrB+, and uvrC+ gene products from Escherichia coli. The uvrB+ and uvrC+ products cochromatograph on DEAE-cellulose and are completely resolved from the uvrA+ product, which has been further purified by phosphocellulose chromatography of the nonadsorbed protein fraction from the DEAE-cellulose. Neither the uvrB+/uvrC+ nor the uvrA+ product shows appreciable endonuclease activity on UV-irradiated DNA when tested separately. However, these factors complement each other to yield and ATP-dependent endonuclease activity specific for UV-irradiated DNA. Gel filtration experiments with the partially purified proteins indicate that the functional uvrA+ gene product has a molecular weight of 100,000. The uvrB+ gene product has an apparent molecular weight of 70,000, but it is presently unclear if this is the size of the uvrB+ product alone or the size of a complex of the uvrB+ and uvrC+ gene products.
摘要
一种体外互补分析方法已被用于从大肠杆菌中部分纯化uvrA+、uvrB+和uvrC+基因产物。uvrB+和uvrC+产物在DEAE-纤维素上共层析，并与uvrA+产物完全分离，uvrA+产物已通过对DEAE-纤维素上未吸附蛋白质部分进行磷酸纤维素层析进一步纯化。单独测试时，uvrB+/uvrC+产物和uvrA+产物对紫外线照射的DNA均未显示出明显的内切核酸酶活性。然而，这些因子相互补充，产生了对紫外线照射的DNA具有特异性的依赖ATP的内切核酸酶活性。对部分纯化蛋白质进行的凝胶过滤实验表明，功能性uvrA+基因产物的分子量为100,000。uvrB+基因产物的表观分子量为70,000，但目前尚不清楚这是否只是uvrB+产物的大小，还是uvrB+和uvrC+基因产物复合物的大小。
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Reconstitution of an Escherichia coli repair endonuclease activity from the separated uvrA+ and uvrB+/uvrC+ gene products.从分离的uvrA⁺和uvrB⁺/uvrC⁺基因产物重建大肠杆菌修复内切核酸酶活性。
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