野桑蚕Hippo通路核心基因的鉴定与序列比较分析.docxVIP

野桑蚕Hippo通路核心基因的鉴定与序列比较分析 孟刚 彭云武 王瑞娴 摘要：Hippo信号通路在动物生长发育过程中起到关键作用。根据已知的Hippo通路组成基因序列，利用BLAST软件同源鉴定野桑蚕（Bombyx mandarina）转录组中的Hippo通路组成基因序列，利用生物信息学工具（ORFinder、web CD-search tool、Expasy-protparam、EMBL-EBI、MEME）分析蛋白质相对分子质量、等电点、二级结构、保守结构域和保守基序等。利用Clustalx和MEGA 6.0进行同源序列比对并构建系统进化树。在野桑蚕中鉴定了salvador、warts、mats、yorkie和scalloped同源基因，并预测了ORF框和编码蛋白。结果表明，野桑蚕Warts、Sav和Mats与家蚕同源蛋白的一致性均在98%以上，Yki和Scalloped与家蚕一致性分别为89.02%和79.13%。比较两者差异，家蚕Yki、Sd蛋白分别缺失了近WW2结构域和在YAP结合结构域中存在蛋白片段插入，野桑蚕Sd蛋白缺失了核定位信号基序和脯氨酸富集基序之间的铰链区。Yki和Sd可按照物种科属分别聚类，野桑蚕与家蚕聚为一支，表明这些同源蛋白在功能相似的同时，也具有基于物种的功能特异性。本研究为深入研究野桑蚕Hippo通路的作用机制提供理論基础。 关键词：野桑蚕（Bombyx mandarina）;Hippo通路;聚类分析 Abstract： The Hippo signaling pathway plays a key role in animal growth and development.According to the known sequences， we used BLAST software to identify the sequence of Hippo pathway component gene in the transcriptome of wild silkworm （Bombyx Mandarina）. The protein molecular weight， isoelectric point， secondary structure， conserved domain and conserved element were also prospected and analyzed by ORFinder， web CD-search tool， Expasy-protparam， EMBL-EBI and MEME bioinformatics tools. The homologous sequence alignment and phylogeny were analyzed by Clustalx and MEGA 6.0 software. The homologous genes of salvador， warts， mats， yorkie and scalloped were identified in wild silkworm， and ORF frame and coding protein were predicted. The result showed that，Warts， Sav and Mats of wild silkworm shared up than 98% similarity with that of domestic silkworm， while Yki and Scalloped shared 89.02% and 79.13% identity respectively between wild and domestic silkworm. The WW2 domain of BmYKi were deleted， and one peptide were inserted in YAP-binding domain of BmSd， while Sd of wild silkworm missing the hinge region between nuclear localization signal motif and proline-rich motif. Yki and Sd could be clustered into subgroup according to species， families and genera， wild and domestic silkworm were clustered into the same branch，suggesting that t