CYR61基因的克隆、表达及系统进化树的构建
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国家高技术研究发展计划(2011AA100401);公益性行业(农业)科研专项(200903045)


Molecular cloning and expression and phylogenetic trees’ construction of CYR61 gene in common carp
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    摘要:

    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。

    Abstract:

    CYR61,CTGF and NOV constitute the CCN family. We used BLASTN program at the NCBI to identify near matches in the common carp (Cyprinus carpio) genome database to each zebrafish (Danio rerio) CYR61 .CYR61 A, CYR61 B, and CYR61 C were identified respectively. ORF of the 3 duplicates of CYR61 were cloned and sequenced in common carp. CYR61 A was 1 158 bp encoding polypeptides of 385 amino acids with a calculated Mw of 42.55 ku and PI of 7.83. CYR61 B was 1 053 bp. CYR61 C was 1 107 bp encoding polypeptides of 368 amino acids with a calculated Mw of 40.83 ku and PI of 8.54. CYR61 A and CYR61 C have 5 exons and 4 introns. CYR61 A, CYR61 B, and CYR61 C all contained 4 conservative domains in common carp. Phylogenetic relationship analysis indicated that CYR61 genes of common carp have higher homology with other species’ CYR61 genes. Differentiation time of CYR61 A, CYR61 B and CYR61 C were earlier than the divergence time between common carp and zebrafish by using MEGA 5.0 software and divergence time formula analysis. RTPCR analysis revealed CYR61 C to be expressed in most common carp tissues, with the principal expression in ovary and weakest expression in heart. CYR61 A was predominantly expressed in testicle and not detected in kidney and body kidney. CYR61 B was highly observed in testicle and brain. Quantitative realtime PCR were used to analyse the expression of CYR61 in common carp development, the result indicated that CYR61 transcriptions expressed highly at the initial stage and lowest at 18 hpf or 24 hpf and then increased before 3d, but decreased at 6 d.

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孙 婷,刘 伟,徐 鹏,孙效文.鲤CYR61基因的克隆、表达及系统进化树的构建[J].上海海洋大学学报,2012,21(3):321-330.
SUN Ting, LIU Wei, XU Peng, SUN Xiao-wen. Molecular cloning and expression and phylogenetic trees’ construction of CYR61 gene in common carp[J]. Journal of Shanghai Ocean University,2012,21(3):321-330.

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  • 在线发布日期: 2012-05-14
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