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通过鲤(Cyprinus carpio)基因组序列与斑马鱼(Danio rerio)CYR61基因编码区全序列的比对得到了CYR61 A、CYR61 B和CYR61 C共3条序列。分别克隆、测序得到其开放阅读框(Open Reading Frame,ORF)分别是1 158 bp、1 053 bp、1 107 bp,CYR61 A和CYR61 C得到完整编码区,都是由5个外显子和4个内含子构成,分别编码385、368个氨基酸,其理论等电点值分别是7.83、8.54,分子量(Mw值)分别为42.55 ku、40.83 ku。鲤3个CYR61基因具有高度同源性,并且均含IB、vWC、TSP1、CT 4个模块。分子系统学分析表明鲤CYR61基因与其他物种CYR61基因具有高度同源性,且CYR61 C与其他高等物种的CYR61同源性更高,利用MEGA 5.0计算出CYR61 A、CYR61 B和CYR61 C的分化时间早于鲤和斑马鱼的物种分化。CYR61 C在13个组织中均有表达,在卵巢中表达最高,肠、脑、鳃次之,在其他组织中表达较低且在心脏中表达最低。CYR61 A在精巢、肠、鳃中表达较高。CYR61 B在精巢和脑中表达最高,肠、肌肉、卵巢次之。实时荧光定量PCR分析CYR61 3个复制在鲤胚胎发育时期的表达,都是在0 h相对表达量最高,显著降低至18 h或24 h,随后逐渐升高并在6 d时下降。
Three sequences of CYR61 A, CYR61 B and CYR61 C were obtained by aligning Cyprinus carpio genomic sequence with the complete coding sequence of CYR61 gene in Danio rerio. Were cloned and sequenced respectively. The ORFs of them were 1 158 bp, 1 053 bp, 1 107 bp, CYR61 A and CYR61 C, respectively, and all of them were composed of 5 exons and 4 Which contains 385,368 amino acids. The theoretical isoelectric point values are 7.83 and 8.54, respectively. The molecular weights (Mw) are 42.55 ku and 40.83 ku, respectively. The 3 CYR61 genes of the common carp were highly homologous and contained 4 modules, IB, vWC, TSP1 and CT. Molecular phylogenetic analysis showed that the CYR61 gene in Cyprinus carpio shared highly homology with CYR61 gene in other species, and CYR61 C had higher homology with CYR61 in other higher species. The differentiation time of CYR61 A, CYR61 B and CYR61 C was calculated using MEGA 5.0 Differentiation of species earlier than common carp and zebrafish. CYR61 C was expressed in all 13 tissues, with the highest expression in ovary, followed by gut, brain and gill, and lower in other tissues and lowest in heart. CYR61 A is highly expressed in the testis, gut and gill. CYR61 B is highest in testes and brains, followed by gut, muscle and ovary. Real-time quantitative PCR analysis of CYR61 3 replication in the common carp embryonic development expression, the highest relative expression at 0 h, significantly reduced to 18 h or 24 h, then gradually increased and decreased at 6 d.