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鲤鱼快肌肌球蛋白重链亚型的cDNA克隆及其与温度驯化相关的基因表达

cDNA cloning of myosin heavy chain isoforms from carp fast skeletal muscle and their gene expression associated with temperature acclimation.

作者信息

Imai J, Hirayama Y, Kikuchi K, Kakinuma M, Watabe S

机构信息

Laboratory of Aquatic Molecular Biology and Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.

出版信息

J Exp Biol. 1997 Jan;200(Pt 1):27-34. doi: 10.1242/jeb.200.1.27.

Abstract

We have isolated cDNA clones encoding fast skeletal muscle myosin heavy chains of carp acclimated to 10, 20 and 30 degrees C for over 5 weeks. All clones covered at least the full length of L-meromyosin, the C-terminal part of the myosin molecule. Nucleotide sequence analysis on cDNA clones showed three types of 3' untranslated sequences, demonstrating that carp expresses at least three myosin heavy chain isoforms in fast skeletal muscle in an acclimation-temperature-dependent manner. cDNAs were identified which were the predominant types expressed in 10 degrees C- and 30 degrees C-acclimated fish, as well as an intermediate type present at all acclimation temperatures. Northern blot analysis using probes of three kinds of DNA fragments from the 3' untranslated region of carp acclimated to 10, 20 and 30 degrees C further confirmed the presence of acclimation-temperature-specific isoforms. In addition, it was found that mRNA levels of three isoforms were altered in an acclimation-temperature-dependent manner. When the deduced amino acid sequences of three types of carp L-meromyosin were compared with those of homoiotherms, the 30 degrees C-acclimated type was more similar to those of homoiotherms than was the 10 degrees C-acclimated type.

摘要

我们已经分离出编码适应10℃、20℃和30℃超过5周的鲤鱼快肌肌球蛋白重链的cDNA克隆。所有克隆至少覆盖了L-肌球蛋白的全长,即肌球蛋白分子的C末端部分。对cDNA克隆的核苷酸序列分析显示出三种类型的3'非翻译序列,表明鲤鱼在快肌中以适应温度依赖的方式表达至少三种肌球蛋白重链异构体。鉴定出了在适应10℃和30℃的鱼中表达的主要类型的cDNA,以及在所有适应温度下都存在的一种中间类型。使用来自适应10℃、20℃和30℃的鲤鱼3'非翻译区的三种DNA片段作为探针进行的Northern印迹分析进一步证实了存在适应温度特异性异构体。此外,还发现三种异构体的mRNA水平以适应温度依赖的方式发生改变。当将三种类型的鲤鱼L-肌球蛋白的推导氨基酸序列与恒温动物的序列进行比较时,适应30℃类型比适应10℃类型与恒温动物的序列更相似。

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