低密度脂蛋白诱导下调的新基因cDNA的克隆及组织表达  

Cloning and tissue expression of a novel gene down-requlated by low density lipoprotein

作　　者：张文成[1] 陈保生[1] 曾武威[1] 吴刚[1] 薛红[1] 

机构地区：[1]中国协和医科大学基础医学研究所医学分子生物学国家重点实验室

出　　处：《基础医学与临床》2003年第3期279-282,共4页Basic and Clinical Medicine

基　　金：国家"九七三"高科技基础研究课题项目 (973 G2 0 0 0 0 5 6 92 0 ) ;中国博士后科学基金

摘　　要：用改进的mRNA差异显示 PCR技术分析ECV30 4在低密度脂蛋白的诱导下表达水平明显差异的cDNA。获得一差异表达的 2 6 5bp新EST ;以该EST为探针 ,从人主动脉cDNA文库中筛选到一个 172 6bp的cDNA克隆 ,其 74 8 12 6 6bp之间的 5 19个碱基构成一个完整的开放阅读框架 ,编码 172个氨基酸组成的蛋白质。其羧基端 94 172区间氨基酸序列中含有三个重复的C2H2型锌指蛋白基序CX2CX3FX5LX2HX3H ,NorthenBlot证实两组ECV30 4中均出现一条 1 7kb的区带 ,LDL诱导后其表达降低了 2 2倍。与差异显示 PCR的结果一致 ,该基因被定名为LRZFG。LRZFG是多组织表达的基因 。Improved Differential display reverse transcription PCR was used to analyze the differentially expressed cDNA in human umbilical vein endothelial cell line ECV304 induced by low density lipoprotein (LDL). A novel differential expressed sequence tag (EST) was obtained. Using this EST as a probe, we screened a full length cDNA of a novel gene's. The full length cDNA is 1726bp. Its 516bp open reading frame run from ATG start codon to a TAA stop codon,encoding a deduced protein sequence of 172 amino acids. Amino acid Sequence analysis showed that its carboxyl terminal contained three repeated tandem C2H2 type zinc fingers protein motif (CX2CX3FX5LX2HX3H).The gene was named as LDL related zinc finger gene (LRZFG). The down regulated expression of LRZFG in ECV304 cells stimulated by LDL was testified by northern blot analysis. LRZFG was expressed in multiple tissues. The function of the novel gene and its role in pathogenesis of atherosclerosis remains to be elucidated.

关 键 词：低密度脂蛋白 MRNA差异显示 基因克隆 CDNA 动脉粥样硬化 

分 类 号：R346[医药卫生—基础医学]