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张敏, 赵金良, 邓燕飞. 2010: 鳜肌酸激酶M-CK cDNA的克隆与组织表达分析. 动物学研究, 31(1): 77-83. DOI: 10.3724/SP.J.1141. 2010.01077
引用本文: 张敏, 赵金良, 邓燕飞. 2010: 鳜肌酸激酶M-CK cDNA的克隆与组织表达分析. 动物学研究, 31(1): 77-83. DOI: 10.3724/SP.J.1141. 2010.01077
ZHANG Min, ZHAO Jin-Liang*, DENG Yan-Fei. 2010. Cloning and Tissue Expression Analysis of Creatine Kinase (M-CK) cDNA from the Mandarin Fish,Siniperca chuatsi. Zoological Research, 31(1): 77-83. DOI: 10.3724/SP.J.1141. 2010.01077
Citation: ZHANG Min, ZHAO Jin-Liang*, DENG Yan-Fei. 2010. Cloning and Tissue Expression Analysis of Creatine Kinase (M-CK) cDNA from the Mandarin Fish,Siniperca chuatsi. Zoological Research, 31(1): 77-83. DOI: 10.3724/SP.J.1141. 2010.01077

鳜肌酸激酶M-CK cDNA的克隆与组织表达分析

Cloning and Tissue Expression Analysis of Creatine Kinase (M-CK) cDNA from the Mandarin Fish,Siniperca chuatsi

  • 摘要: 利用RT–PCR和cDNA末端快速扩增法(RACE)克隆了鳜(Siniperca chuatsi)肌酸激酶(creatine kinase,CK)cDNA序列,并分析了该基因的结构特征和系统关系。鳜CK cDNA序列全长1586 bp,包括5′端非翻译区92 bp,3′端非翻译区348 bp和开放阅读框(ORF)1 146 bp,共编码381个氨基酸。鳜CK具有脊椎动物CK共有的保守结构域和肌型肌酸激酶(M-CK)同工酶的特异识别位点;氨基酸序列与M-CK型的相似度最高,而与脑型肌酸激酶(B-CK)和线粒体型肌酸激酶(Mi-CKs)的相似度较低;在CK系统关系树中鳜CK与M-CK群聚类。这些均表明,鳜CK属脊椎动物M-CK型。RT-PCR分析表明,鳜M-CK在成体不同组织中的表达量不同,其中,在皮肤、卵巢、肾脏、胃、肌肉和心脏中表达较强;而在眼和脑、肝胰脏中表达较弱。

     

    Abstract: The creatine kinase(CK) cDNA from the mandarin fish Siniperca chuatsi was cloned by RT-PCR and rapid amplification of cDNA ends (RACE) methods. The structural characteristics and phylogeny of this gene were analyzed. Sequence analysis revealed a 1 586 bp cDNA sequence containing 92 bp 5′-untranslated region, 348 bp 3′-untranslated region and 1146 bp open reading frame (ORF), which encoded 381 amino acids. Conserved sequence blocks of vertebrate CKs and diagnostic boxes for the muscle CK(M-CK) isozyme were identified in S. chuatsi CK. Siniperca chuatsi CK showed a higher similarity with vertebrates M-CK isozyme than other CK isozymes (Brain CK, Mitochondrial CKs) and grouped with M-CK isozyme in CK phylogeny, which strongly supported that S. chuatsi CK belongs to M-CK isozyme type. Semi-quantitative RT-PCR analysis demonstrated that the M-CK transcript expression varied among the different tissues and was detected at a high level in skin, ovary, kidney, stomach, muscle and heart, but lower in eye, brain and liver.

     

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