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。
-
关键词:
- 鳜 /
- 肌酸激酶 /
- cDNA末端快速扩增 /
- 组织表达
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. -
点击查看大图
计量
- 文章访问数: 4037
- HTML全文浏览量: 248
- PDF下载量: 1877
- 被引次数: 0
下载:
