HDAC2 cDNA 的克隆及在酵母细胞中的表达

doi:10.3969/j.issn.1004-616x.2000.03.001

癌变·畸变·突变 ›› 2000, Vol. 12 ›› Issue (3): 129-132.doi: 10.3969/j.issn.1004-616x.2000.03.001

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HDAC2 cDNA 的克隆及在酵母细胞中的表达

陈　坚1 ,张晓琴2 , 傅继梁3

11 第二军医大学海军卫生教研室,上海　200433 ; 21 第二军医大学长海医院感染科,上海　200433 ;31 第二军医大学医学生物技术和分子遗传研究所,上海　200433

收稿日期:2000-01-13 修回日期:2000-02-25 出版日期:2000-07-30 发布日期:2000-07-30

CLONING OF THE HISTONE DEACETYLASE 2 cDNA AND IT’S EXPRESSION IN YEAST

CHEN Jian1 , ZHAN G Xiao-qin2 , FU Ji-liang3

1. Department of Naval Hygiene , 2. Inf ection Department Changhai Hospital , 3. Instit ute of Medical Biotechnology Molecular Genetics , Second Military Medical University , S hanghai , 200433 , China

Received:2000-01-13 Revised:2000-02-25 Online:2000-07-30 Published:2000-07-30

摘要/Abstract

摘要： 目的:本研究的目的为获得HDAC2 的cDNA 及构建酵母双杂交系统中的靶基因。方法:利用RT - PCR 方法,从人HeLa 细胞中扩增出一约115Kb 的DNA 片段,全自动测序后,重组入pLexA 载体,构建成pLexA - HDAC2 ,用醋酸锂法转化酵母菌EGY48 (p8op - lacZ) 。结果:结果显示,获得的1. 5Kb 的DNA 片段碱基序列与文献报道的HDAC2 的cDNA 一致。pLexA - HDAC2 转化EGY48 (p8op - LacZ) 3 天后,长出1mm 大小的白色菌落。结论:上述结果表明pLexA - HDAC2 可在EGY48(p8op - LacZ) 稳定表达,无毒性,也没有自身激活LacZ的功能,可作为酵母双杂交系统中的靶基因。

关键词: 组蛋白脱乙酰化, 组蛋白脱乙酰化酶2, 酵母双杂交系统

Abstract: Purpose : In order to obtain histone deacetylase 2 (HDAC2) cDNA and const ruct the target gene of yeast two hybrid system. Methods : we amplified about 1. 5 Kb DNA f ragment f rom HeLa cell using RT - PCR method. After automatic sequeced , a recombinant plasmid pLexA - HDAC2 was made and t ransformed into yeast EGY48 (p8op - Lac2) , and the expression of pLexA - HDAC2 in the yeast was observed. Results : Results show that : The sequence of RT - PCR product was same with the sequence of reported HDAC2 cDNA. About 1mm white yeast clone was grown in the selective medium after 3 days. Conclusion : The result s indicated that : HDAC2 cDNA has been obtained. pLexA - HDAC2 can be expressed in yeast with inactivation of LacZ gene and nontoxic to yeast , and can be as a target gene of yeast two hybrid system.

Key words: histone deacetylation, histone deacetylase2 (HDAC2), yeast two hybrid system

中图分类号:

Q784

陈　坚, 张晓琴, 傅继梁. HDAC2 cDNA 的克隆及在酵母细胞中的表达[J]. 癌变·畸变·突变, 2000, 12(3): 129-132.

CHEN Jian , ZHAN G Xiao-qin , FU Ji-liang. CLONING OF THE HISTONE DEACETYLASE 2 cDNA AND IT’S EXPRESSION IN YEAST[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2000, 12(3): 129-132.

HDAC2 cDNA 的克隆及在酵母细胞中的表达

CLONING OF THE HISTONE DEACETYLASE 2 cDNA AND IT’S EXPRESSION IN YEAST

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