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孔庆辉,晁燕,夏明哲,李长忠,祁得林.2016.黄河裸裂尻鱼肌酸激酶M-CK cDNA的克隆及组织表达分析.动物学杂志,51(1):84-94.
黄河裸裂尻鱼肌酸激酶M-CK cDNA的克隆及组织表达分析
cDNA Cloning and Tissue Expression Analyses of Creatine Kinase Gene from Schizopygopsis pylzovi
投稿时间:2015-06-26  修订日期:2015-12-29
DOI:DOI: 10.13859/j.cjz.201601010
中文关键词:  黄河裸裂尻鱼  肌酸激酶  基因克隆  表达
英文关键词:Schizopygopsis pylzovi  Creatine kinase  Gene cloning  Expression
基金项目:国家自然科学基金项目(面上项目,重点项目,重大项目)
作者单位E-mail
孔庆辉 青海大学农牧学院动物科学系 西宁 kc1008611@163.com 
晁燕 青海大学农牧学院动物科学系 西宁  
夏明哲 青海大学农牧学院动物科学系 西宁  
李长忠 青海大学生态环境工程学院 西宁  
祁得林 青海大学农牧学院动物科学系 西宁 delinqi@126.com 
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中文摘要:
      肌酸激酶(Creatine kinase, CK)能够催化在二磷酸腺苷ADP和磷酸肌酸之间磷酸基的可逆性转移,在细胞能量代谢过程中发挥重要作用。本研究利用RT-PCR和RACE方法克隆了黄河裸裂尻鱼(Schizopygopsis pylzovi)肌酸激酶基因全长cDNA序列,并进行了生物信息学分析和系统发育关系研究。结果表明,黄河裸裂尻鱼肌酸激酶cDNA全长为1599 bp,开放阅读框(ORF)长1143bp,编码380个氨基酸,具有典型的N-端结构域和C-端催化结构域。氨基酸序列同源性分析表明,黄河裸裂尻鱼肌酸激酶与斑马鱼和鲤M3-CK有较高的同源性,其序列一致度达94%以上。系统发育分析显示,黄河裸裂尻鱼肌酸激酶与斑马鱼和鲤的M3-CK聚在一支,形成一个单系群,初步判定克隆获得的黄河裸裂尻鱼肌酸激酶基因可能是鲤和斑马鱼M3-CK的直系同源基因。利用Real-time PCR方法检测和分析了主要组织肌酸激酶mRNA表达水平,结果表明黄河裸裂尻鱼肌肉、肠、眼、心脏中肌酸激酶转录本表达较高,在肝胰脏和脑中表达微弱。肌酸激酶在肌肉、肠、和心脏中高表达与其能量代谢功能相适应,而在眼组织中的高表达是否与肌酸激酶的其他功能相关,有待于进一步研究。
英文摘要:
      Creatine kinase(CK)plays an important role in the process of cell energy metabolism by catalyzing the reversible transfer of phosphate group between ADP and phosphocreatine. The cDNA full-length of the creatine kinase gene from Schizopygopisis pylzovi was cloned using RT-PCR and RACE methods, and then conducted the bioinformatics analyses. The results showed that the full-length cDNA of CK gene was 1599 bp, including a 1143 bp complete ORF encoding 380 amino acid peptide. The creatine kinase of Schizopygopsis pylzovi contains a N-terminal domain and a C-terminal catalysis domain. Sequence homologous analysis of amino acids showed that the CK of Schizopygopsis pylzovi has high homology with the M3-CK of Danio rerio and Cyprinus carpio, the identity reaches above 94%. Phylogenetic analysis showed that the CK of Schizopygopsis pylzovi and the M3-CK of Danio rerio and Cyprinus carpio were clustered together and formed a monophyletic group, from which the CK of Schizopygopsis pylzovi could be tentatively identified as the orthologous gene of the M3-CK of Danio rerio and Cyprinus carpio. The mRNA levels of major tissues of Schizopygopsis pylzovi were detected and analyzed using Quantitative Real-time PCR. The result showed that the CK transcript was highly expressed in muscle, intestine, eye and heart, while weakly expressed in hepatopancreas and brain. The high expression in muscle, intestine and heart of CK is consistent with its function in cell energy metabolism, but the high expression in eyes may be related to other functions of CK, need to further researches.
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