不同原沉默子对基因沉默的作用

J4 ›› 2011, Vol. 37 ›› Issue (2): 241-244.

不同原沉默子对基因沉默的作用

张新民|刘楠|于群|王浩天|颜炜群|毕鑫

1.吉林大学药学院生物工程教研室|吉林长春 130021；2.美国罗彻斯特大学生物学系|美国纽约 14627

收稿日期:2010-12-08 出版日期:2011-03-28 发布日期:2011-03-28
通讯作者: 颜炜群(Tel：0431-85619716，E-mail：weiqunyanjlu@126.com) E-mail:weiqunyanjlu@126.com
作者简介: 张新民（1983-）|男|甘肃省临洮市人|在读医学博士|主要从事分子生物学方面的研究。
基金资助:
美国国立卫生院基金项目资助课题(GM68484)；中国留学基金委员会建设高水平大学项目资助课题(2008)

Effects of different protosilencers on gene silencing

ZHANG Xin-min^1,2,LIU Nan¹,YU Qun²,WANG Hao-tian¹,YAN Wei-qun ¹,BI Xin²

1.Department of Biological Engineering, School of Pharmacy, Jilin University,Changchun 130021,China；2.Department of |Biology,Rochester University,New York |14627,USA

Received:2010-12-08 Online:2011-03-28 Published:2011-03-28

摘要/Abstract

摘要：

的：探讨不同原沉默子对基因沉默的作用和基因沉默差异的原因。方法：用不同原沉默子替换HML-I沉默子，在有无HML-E沉默子时，比较报告基因URA3表达量的变化以及染色质结构的不同。结果：删除HML-E沉默子且HML-I替换成原沉默子RAP1p、ORC和ABF1结合位点的3株酵母菌种在SC-ura 及SC+FOA平板上的生长状态相同；HML-E沉默子存在的条件下，HML-I替换成原沉默子RAP1p、ORC和ABF1结合位点的3株酵母菌种在SC-ura平板上的生长状态没有区别；在FOA平板上，HML-I替换成原沉默子RAP1p结合位点的菌株无法生长，HML-I替换成ABF1结合位点以及ORC结合位点的菌株能够存活并形成了与HML-I替换成HMR-E菌株相似的特异性泳带。结论：沉默子与原沉默子相互作用对基因表达起一定程度的沉默作用，染色质结构的差异可能导致了基因沉默的差异。

关键词: 基因沉默；沉默子；原沉默子；微球菌核酸酶

Abstract:

Abstract:Objective To study the effects of different protosilencers on gene silencing and the reasons of differences of gene silencing. Methods HML-I silencer was replaced by different protosilencers. The expression level of reporter gene and the structure of chromosome were compared with or without HML-E. Results After deleting HML-E silencer and replacing HML-I with RAP1p,ORC and Abflp respectively,the cells grew similarly inSC-ura and SC+FOA plate. When HML-E exited,the cells grew similarly on SC-ura plate after replacing HML-I with RAP1p,ORC and ABF1 respectively. The cells were unable to grow on FOA plate when HML-I was replaced with Raplp,However,the cells grew on FOA plate when HML-I was replaced with ORC or ABF1.The micrococcal nuclease digestion data showed that the cells which HML-I was replaced with ORC or ABF1presented similar specific bands to which HML-I was replaced by HMR-E. Conclusion Protosilencers can’t silence gene in activated and basal expression independently,but it can do it with the help of silencer protosilencer. The differences in chromatin structure might be the reason for the difference of gene silencing.

Key words: gene silencing;silencer;protosilencer;micrococcal nuclease

中图分类号:

Q342.3

张新民|刘楠|于群|王浩天|颜炜群|毕鑫. 不同原沉默子对基因沉默的作用[J]. J4, 2011, 37(2): 241-244.