| 引用本文： |
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张雪松,李德凤,薛姜珊,陈孜孟,李卓,蒙健宗.重组Pol D DNA聚合酶的纯化及其功能初步研究[J].广西科学,2022,29(6):1086-1093. [点击复制]
- ZHANG Xuesong,LI Defeng,XUE Jiangshan,CHEN Zimeng,LI Zhuo,MENG Jianzong.Primary Study on Purification and Function of Recombinant Pol D DNA Polymerase[J].Guangxi Sciences,2022,29(6):1086-1093. [点击复制]
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| 摘要: |
| 为研究古菌Pol D DNA聚合酶两个亚基的功能及其相互作用，对重组超嗜热硫还原古菌Thermococcus sp.4557 Pol D DNA聚合酶小亚基DP1和大亚基DP2进行柱层析纯化，采用荧光标记核酸结合聚丙烯酰胺凝胶电泳实验对两个亚基体外功能进行研究。结果表明，DP2具有DNA复制延伸活性，DP1具有3'→5'外切核酸酶活性；DP1在与DP2的浓度比高于0.3∶1时可降解DP2的延伸产物；DP2对DP1的外切核酸酶活性有促进作用，Sld5和Psf1蛋白复合体(Sld5 and Psf1 Complex，GINS 51)对DP1的外切核酸酶活性有抑制作用。古菌Pol D DNA聚合酶大小亚基的功能及其相互作用对DNA复制机制的进一步认识有借鉴意义。 |
| 关键词: 古菌 Pol D DNA聚合酶 亚基 延伸活性 外切酶活性 |
| DOI：10.13656/j.cnki.gxkx.20230110.008 |
| 投稿时间：2021-02-23修订日期：2021-03-26 |
| 基金项目:十三五海洋经济创新发展示范项目(海洋工具酶的开发与产业化子课题)资助。 |
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| Primary Study on Purification and Function of Recombinant Pol D DNA Polymerase |
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ZHANG Xuesong1, LI Defeng1, XUE Jiangshan1, CHEN Zimeng2, LI Zhuo2, MENG Jianzong3
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| (1.College of Life Science ＆ Technology, Guangxi University, Nanning, Guangxi, 530005, China;2.Third Institute of Oceanography, Ministry of Natural Resources, Xiamen, Fujian, 361000, China;3.Institute of Food ＆ Pharmaceutical Science, Guangxi Agricultural Vocational University, Nanning, Guangxi, 530007, China) |
| Abstract: |
| In order to study the function and interaction of the two subunits of Archaeal Pol D DNA polymerase,the small subunit DP1 and large subunit DP2 of recombinant hyperthermophilic archaeal Thermococcus sp.4557 Pol D DNA polymerase were purified by column chromatography.The function of the two subunits in vitro was studied by fluorescence labeling nucleic acid combined with polyacrylamide gel electrophoresis experiments.The results showed that DP2 had DNA replication elongation activity,and DP1 had 3'→5' exonuclease activity.When the concentration ratio of DP1 to DP2 is higher than 0.3∶1,the extended product of DP2 can be degraded.DP2 promoted the exonuclease activity of DP1,while Sld5 and Psf1 Complex (GINS 51) had an inhibitory effect on DP1 exonuclease activity.The function and interaction of the large and small subunits of Archaeal Pol D DNA polymerase have reference significance for further understanding of DNA replication mechanism. |
| Key words: Archaea Pol D DNA polymerase subunit extension activity exonuclease activity |
