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在体内骨骼肌发育过程中,使用一种被鉴定为编码小鼠骨骼肌成年快速肌球蛋白重链的重组质粒检测到编码不同肌球蛋白重链亚型的mRNA的顺序积累。

Sequential accumulation of mRNAs encoding different myosin heavy chain isoforms during skeletal muscle development in vivo detected with a recombinant plasmid identified as coding for an adult fast myosin heavy chain from mouse skeletal muscle.

作者信息

Weydert A, Daubas P, Caravatti M, Minty A, Bugaisky G, Cohen A, Robert B, Buckingham M

出版信息

J Biol Chem. 1983 Nov 25;258(22):13867-74.

PMID:6196357
Abstract

In order to study developmental transitions of myosin heavy chain gene expression, we have cloned from newborn mouse skeletal muscle a recombinant plasmid (plasmid MHC 32) that contains an insertion coding for the COOH-terminal portion of an adult fast myosin heavy chain isoform of mouse skeletal muscle. By Northern blots and dot blots, it has been shown that the MHC 32 sequence reveals a broad cross-hybridization with RNA from different mammalian striated muscle tissues. Southern blots with mouse genomic DNA show only one homologous gene, but cross-hybridization at lower stringency to seven to eight different bands, some containing multiple genomic fragments, among which are probably the genes encoding the different striated muscle isoforms. S1 protection experiments with RNA from mouse skeletal muscle before and after birth demonstrate that plasmid MHC 32 is homologous to a major mRNA species of adult skeletal muscle. This adult mRNA is a predominant sequence within 5-6 days after birth. It begins to accumulate at 1-3 days; at the 18th day fetal stage, another major mRNA species is detected as partially homologous with the adult MHC 32 sequence. This fetal myosin heavy chain mRNA is still predominant at 1-3 days after birth, but is rapidly (by 5-6 days) replaced by the adult MHC sequence. There is thus a rapid transition after birth from fetal to adult skeletal muscle myosin heavy chain mRNA sequences.

摘要

为了研究肌球蛋白重链基因表达的发育转变,我们从小鼠新生骨骼肌中克隆了一个重组质粒(质粒MHC 32),该质粒含有一段编码小鼠骨骼肌成年快速肌球蛋白重链同工型COOH末端部分的插入片段。通过Northern印迹和斑点印迹表明,MHC 32序列与来自不同哺乳动物横纹肌组织的RNA呈现广泛的交叉杂交。用小鼠基因组DNA进行的Southern印迹显示只有一个同源基因,但在较低严谨度下与7至8条不同条带交叉杂交,其中一些包含多个基因组片段,其中可能是编码不同横纹肌同工型的基因。对出生前后小鼠骨骼肌RNA进行的S1保护实验表明,质粒MHC 32与成年骨骼肌的一种主要mRNA种类同源。这种成年mRNA在出生后5 - 6天内是主要序列。它在出生后1 - 3天开始积累;在胎儿期第18天,检测到另一种主要mRNA种类与成年MHC 32序列部分同源。这种胎儿肌球蛋白重链mRNA在出生后1 - 3天仍然占主导,但在出生后5 - 6天迅速被成年MHC序列取代。因此,出生后从胎儿骨骼肌到成年骨骼肌的肌球蛋白重链mRNA序列存在快速转变。

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