应用三元递减法筛选特异性心脏生长相关基因

Identification of the Specifically Cardiac Growth Related Gene by Three Elements PCR-select cDNA Subtraction

心脏是由胚胎干细胞特异性分化而来的 ,但其分化的分子生物学机制尚不十分了解 .为建立一种新的筛选特异性心脏相关基因的方法 ,克隆特异性心脏生长相关基因 .从胚胎心、成年心和去胎心的胚胎中提取 m RNA,建立胎心 /成年心和胎心 /胎身两个递减性 c DNA文库 ,通过 DNA芯片和微阵列杂交筛选和克隆 ,建立了三元递减克隆的新方法 .获得了一个全长为 1 0 0 6 bp可编码1 94个氨基酸的新的与心脏生长相关的基因 ,它是 LIM家族的新成员 ,可特异性在心肌细胞表达 ,并可促进心肌细胞的生长 .结果表明 ,三元递减筛选法可以应用于寻找新的组织特异性表达的基因 .并且获得了一个新的与心脏生长相关的新基因 。

Heart is specifically derived from embryonic stem cells.The molecular mechanism of heart differentiation,however,still remains undefined.To develop a new means for searching genes specifically related to heart and to clone genes specific to heart growth,the mRNAs was isolated from embryonic heart,mature heart,and embryotic body without heart and established two subtractive cDNA libraries (embryonic heart/mature heart and embryonic heart/embryonic body without heart).Utilizing DNA chips and microarray hybridization selection,a novel method three element subtraction was established.Using this new method,a new heart related gene was obtained.The full length of it was 1 006 bp and it encoded a protein with 194 amino acids.It is a new member of Lim gene family.It can specifically express in cardiac muscle cells and can promote the growth of cardiac myocytes.Taken together,the results indicate that the three element subtraction method can be used to search novel tissue specific genes and a new gene related to the growth of heart was obtained.The gene may plays an important role in the growth of cardiac myocytes and may be involved in the pathogenesis of hypertrophy.