中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (10): 1773-1776.doi: 10.3969/j.issn.1673-8225.2011.10.014

• 干细胞转基因表达 transgenic expression in stem cells • 上一篇    下一篇

起始识别复合物结合位点对沉默子的辅助作用

张新民1,2，王浩天1，刘  楠1，于  群2，颜炜群1，毕  鑫2   

  1. 1吉林大学药学院，吉林省长春市  130021；2罗彻斯特大学生物系，纽约州罗彻斯特，美国  14627
  • 收稿日期:2010-12-01 修回日期:2011-01-25 出版日期:2011-03-05 发布日期:2011-03-05
  • 通讯作者: 颜炜群，教授，吉林大学药学院，吉林省长春市 130021 weiqunyanjlu@126.com
  • 作者简介:张新民☆，男，1983年生，甘肃省临洮县人，汉族，吉林大学医学生物工程学在读博士。 xmzhangjlu@126.com

Auxiliary role of origin recognition complex binding site for silencer involved in transcriptional silencing

Zhang Xin-min1,2, Wang Hao-tian1, Liu Nan1, Yu Qun2, Yan Wei-qun1, Bi Xin2   

  1. 1School of Pharmaceutical Sciences, Jilin University, Changchun  130021, Jilin Province, China; 2Department of Biology, University of Rochester, Rochester, NY  14627, USA
  • Received:2010-12-01 Revised:2011-01-25 Online:2011-03-05 Published:2011-03-05
  • Contact: Yan Wei-qun, Professor, School of Pharmaceutical Sciences, Jilin University, Changchun 130021, Jilin Province, China weiqunyanjlu@126.com
  • About author:Zhang Xin-min☆, Studying for doctorate, School of Pharmaceutical Sciences, Jilin University, Changchun 130021, Jilin Province, China xmzhangjlu@126.com

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摘要：

背景：沉默子由起始识别复合物结合位点，阻遏活化蛋白RAP1等元件组成，人们对于沉默子的功能及相互关系有较多研究，但对组成沉默子的元件与基因沉默之间关系的研究很少。
目的：研究起始识别复合物结合位点对沉默子基因沉默的辅助作用。
方法：用右侧带有报告基因URA3的起始识别复合物结合位点，以及不带任何结合位点的序列替代酵母基因组HML区域的HML-I沉默子后，比较URA3基因表达量，并使用DNA拓扑法分析染色质的结构变化。
结果与结论：HML-E沉默子单独存在的情况下，酵母无法在FOA平板上生长，但在起始识别复合物结合位点的辅助下，能够在FOA平板上生长， 两者之间区域的染色质大多为负超螺旋。因此可知起始识别复合物结合位点能增加HML-E沉默子作用范围，产生这种作用的原因是两者相互作用下形成紧密的染色质结构。

关键词: 沉默子, 起始识别复合物结合位点, 拓扑结构, 酿酒酵母, 基因沉默

Abstract:

BACKGROUND: Silencer is consisted of origin recognition complex (ORC), RAP1 and other elements. There were a lot of reports on function and relationship between silencers but few researches concerning silencer elements.
OBJECTIVE: To study the auxiliary gene silencing effect of ORC to silencer.
METHODS: Delete or replace HML-I silencer with ORC binding site with URA3 in the right side or blank sequence without any binding sites to compare expression level of URA3 and analyze the difference of chromatin structure with DNA topological method.
RESULTS AND CONCLUSION: When HML-E silencer exited alone, yeast could not grow on FOA plates. But with auxiliary effect of ORC binding site, it was able to grow on the FOA plate, and the proportion of negative super coiled DNA was larger compared with that in the case that HML-E silencer exits alone. Therefore, ORC can interact with HML-E to condense the chromatin structure and enhance gene silencing.

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