LENS FIBER DIFFERENTIATION CORRELATED WITH ACTIVATION OF 2 DIFFERENT DNASES IN LENS EMBRYONIC-CELLS
LENS FIBER DIFFERENTIATION CORRELATED WITH ACTIVATION OF 2 DIFFERENT DNASES IN LENS EMBRYONIC-CELLS
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DOI：
10.1016/0922-3371(89)90743-0
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发表时间：
1989-07-01
期刊：
影响因子：
--
通讯作者：
ALLINQUANT, B
中科院分区：
文献类型：
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作者：
COUNIS, MF;CHAUDUN, E;ALLINQUANT, B
为了鉴定晶状体分化组织中存在的不同dna酶，我们使用了一种检测方法，在SDS聚丙烯酰胺凝胶电泳后直接在含dna的凝胶上显示它们的活性。与纯化的微球菌核酸酶相比，在0.1% SDS聚丙烯酰胺凝胶电泳分离后，核胚胎晶体提取物的dna酶不发生复性。当运行缓冲液和分离凝胶中的SDS浓度降低至0.075%时，观察晶状体dna酶的恢复情况和酶活性。等电聚焦在聚丙烯酰胺凝胶中进行，该凝胶覆盖有含有DNA的琼脂糖凝胶，允许可视化DNA酶活性的pI。在11天胚胎晶状体中证实了几种dna酶同工酶的存在。在上皮晶状体核中，具有碱性pI的高分子量同工酶占优势。在有丝分裂后的纤维透镜核中，检测到两个具有酸性pI的低分子量同工酶以及具有碱性pI的高分子量同工酶。
In order to identify the different DNAases present in the lens differentiating tissue, we have used an assay which reveals their activity directly on DNA-containing gels after SDS polyacrylamide gel electrophoresis. DNAase renaturation from nuclear embryonic lens extracts does not occur after separation in 0.1% SDS polyacrylamide gel electrophoresis in contrast to that observed with purified micrococcal nuclease. When the SDS concentration in the running buffer and separating gel is decreased to 0.075%, renaturation of lens DNAase and enzyme activities are observed. Isoelectrofocusing was carried out in a polyacrylamide gel which was overlaid with an agarose gel containing DNA, permitting the visualization of the pI of DNAase activity. The presence of several DNAase isoenzymes was demonstrates in 11-day embryonic lenses. In epithelial lens nuclei, high molecular weight (MW) isoenzymes with basic pI were predominant. In post-mitotic fiber lens nuclei, two lower MW isoenzymes with acidic pI were detected as well as a high MW activity with a basic pI.