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肌酸激酶(CK)能够催化磷酸基在二磷酸腺苷(ADP)和磷酸肌酸之间的可逆性转移,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c DNA序列,并进行了生物信息学分析和系统发育关系研究。黄河裸裂尻鱼肌酸激酶c DNA全长1 599 bp,开放阅读框(ORF)1 143 bp,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(Danio rerio)和鲤(Cyprinus carpio)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与鲤和斑马鱼M3-CK是直系同源基因。利用Real-time PCR方法检测和分析了黄河裸裂尻鱼主要组织肌酸激酶m RNA表达水平,肌肉、肠、眼、心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。
Creatine kinase (CK) catalyzes the reversible transfer of phosphate groups between adenosine diphosphate (ADP) and creatine phosphate and plays an important role in cellular energy metabolism. In this study, the full-length c DNA sequence of creatine kinase gene of Schizopygopsis pylzovi was cloned by RT-PCR and RACE, and the relationship between bioinformatics analysis and phylogenetic relationship was studied. The full length cDNA of creatine kinase c in the Yellow River naked shrimps was 1 599 bp in length and 1 143 bp in open reading frame (ORF) encoding 380 amino acids with typical N-terminal and C-terminal catalytic domains. The amino acid sequence homology analysis showed that the homologous sequences of creatine kinase of Danio rerio and Cyprinus carpio were higher than that of M3-CK of Danio rerio and Cyprinus carpio, and the sequence identity was above 94%. Phylogenetic analysis showed that the M3-CK of the naked nudibranch of the Yellow River was clustered with M3-CK of zebra fish and common carp to form a monophyletic group. It was initially determined that the creatine kinase gene of the naked nudibranch from the Yellow River might be related to the common carp And zebrafish M3-CK are orthologous genes. Real-time PCR method was used to detect and analyze the mRNA expression levels of creatine kinase m RNA in the main tissues of the naked split mackerel in the Yellow River. The expression of creatine kinase in muscle, intestine, eye and heart was higher than that in the hepatopancreas and brain . The high expression of creatine kinase in muscle, intestine and heart adapts to its energy metabolism function. Whether high expression in eye tissue is related to other functions of creatine kinase remains to be further studied.