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大鼠心肌肌球蛋白轻链-2基因的结构、组织与表达。鉴定出一段赋予心脏特异性表达的250个碱基对的片段。

Structure, organization, and expression of the rat cardiac myosin light chain-2 gene. Identification of a 250-base pair fragment which confers cardiac-specific expression.

作者信息

Henderson S A, Spencer M, Sen A, Kumar C, Siddiqui M A, Chien K R

机构信息

Department of Medicine, University of California, San Diego, School of Medicine, La Jolla 92093.

出版信息

J Biol Chem. 1989 Oct 25;264(30):18142-8.

PMID:2808370
Abstract

The present study characterized the structure, organization, and expression of the rat cardiac myosin light chain (MLC) -2 gene. The rat cardiac MLC-2 gene has seven exons which display complete conservation with the exon structure of the rat fast twitch skeletal MLC-2 gene. A 250-base pair (bp) sequence of the 5'-flanking region contains CArG motifs and additional cis elements, each greater than 10 bp in length, which were conserved in sequence and relative position with the chick cardiac MLC-2 gene. A series of MLC-2/luciferase fusion genes consisting of nested 5' deletions of the MLC-2 5'-flanking region were constructed and transfected into primary neonatal rat myocardial cells and a non-myocardial cell line (CV-1), demonstrating that this 250 bp of the MLC-2 5'-flanking region was sufficient to confer cardiac specific expression on a luciferase reporter gene. This study suggests the presence of important proximal regulatory sequences in the MLC-2 5'-flanking region which are capable of directing the cardiac specific expression of the rat cardiac myosin light chain-2 gene.

摘要

本研究对大鼠心肌肌球蛋白轻链(MLC)-2基因的结构、组织和表达进行了表征。大鼠心肌MLC-2基因有七个外显子,其与大鼠快肌骨骼肌MLC-2基因的外显子结构完全保守。5'侧翼区的一段250个碱基对(bp)的序列包含CArG基序和其他顺式元件,每个元件长度大于10 bp,其在序列和相对位置上与鸡心肌MLC-2基因保守。构建了一系列由MLC-2 5'侧翼区嵌套5'缺失组成的MLC-2/荧光素酶融合基因,并将其转染到原代新生大鼠心肌细胞和非心肌细胞系(CV-1)中,表明MLC-2 5'侧翼区的这250 bp足以赋予荧光素酶报告基因心脏特异性表达。本研究表明在MLC-2 5'侧翼区存在重要的近端调控序列,其能够指导大鼠心肌肌球蛋白轻链-2基因的心脏特异性表达。

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