家蚕JAK/STAT信号通路相关基因克隆与分析  

Cloning and Sequence Analysis of JAK/STAT Pathway Related Major Genes of Silkworm (Bombyx mori)

作　　者：彭景琨[1] 曹广力[1] 钱莹[1] 张晓丽[1] 马焕艳[1] 何蕾[1] 薛仁宇[1] 贡成良[1] 

机构地区：[1]苏州大学基础医学与生物科学学院,江苏苏州215123

出　　处：《中国农业科学》2012年第17期3592-3601,共10页Scientia Agricultura Sinica

基　　金：高等学校博士学科点专项科研基金项目(优先发展领域)(20113201130002);国家重点基础研究发展计划"973计划"(2012CB114600);江苏省自然科学基金项目(BK2009117);江苏高校优势学科建设工程项目

摘　　要：【目的】鉴定家蚕JAK/STAT信号通路组成原件,并预测家蚕JAK/STAT信号通路主要相关基因的功能。【方法】在电子克隆的基础上,通过RT-PCR获得家蚕JAK/STAT信号通路的主要相关基因;利用在线软件对相关蛋白进行结构域分析;基于microarray数据库或qPCR分析各基因的组织表达特征。【结果】克隆了家蚕JAK/STAT信号通路主要相关基因,包括BmSTAT、BmHOP、BmSOCS2、BmSOCS5A、BmSOCS5B、BmSOCS6、BmDRK、Bmken、BmPIAS1、BmPIAS2以及BmPI3K,但没有能克隆到果蝇Upd1、Upd2和Upd3的同源体基因。序列分析显示BmHOP具CRD_FZ、Kringle、转膜区域和PKc超家族蛋白激酶催化结构域TyrKc;BmSTAT有2种亚型,均具SH2、STAT_bind、STAT_int和STAT_alpha结构域;在克隆的BmSOCS家族的3个基因中,BmSOCS2A和BmSOCS6A具典型的SH2和SOCS结构域;BmDRK由SH3和SH2两种结构域形成了SH3-SH2-SH3串联结构;BmKen存在BTB和锌指结构域,BmPIAS1和BmPIAS2具有LCR结构域,BmPI3K具有2个SH2结构域。【结论】成功克隆了家蚕JAK/STAT信号通路主要相关基因。相关蛋白的保守结构域分析显示,在家蚕JAK/STAT信号通路中,BmHOP是JAK激酶的一种,可以磷酸化BmSTAT转录因子,激活靶基因转录;BmSOCS、Bmken、BmPIAS1以及BmPIAS2对JAK/STAT通路起负反馈作用。[ Objective ] The objective of this study is to identify the elements of JAK/STAT pathway of silkworm, Bombyx mori and determine the function of JAK/STAT pathway related major genes in silkworm. [Method] JAK/STAT pathway related major genes in silkworm were obtained through electronic cloning and conventional RT-PCR. Domain analysis on the related protein were conducted by using bio-softwares online, and the expression profile of these genes in diverse tissues were analysed through microarray database or qPCR. [Result] The silkworm JAK/STAT pathway related major genes including BmSTAT, BmHOP, BmSOCS2, BmSOCS5A, BmSOCS5B, BmSOCS6, BmDRK, Bmken, BmPIAS1, BmPIAS2 and BmPI3K were cloned, respectively, but the homologues of Drosophila Updl, Upd2 and Upd3 were not cloned. Sequence analysis showed that BmHOP contained CRD FZ, Kringle domains, transmembrane region and Pkc superfamily protein kinase catalysis domain TyrKc. BmSTAT had two isoforms, which were consisted of SH2, STAT_bind, STAT_int and STAT_alpha domains. Both BmSOCS2A and BmSOCS6A had a conserved SH2 domain and a SOCS box in cloned three genes of BmSOCS family. BmDRK was consisted of SH3 and SH2 domains that connected in series to form SH3-SH2-SH3. BmKen contained BTB and zinc finger domains. BmPIAS 1 and BmPIAS2 contained LCR domains. BmPI3K had two conserved SH2 domains. [Conclusion] The JAK/STAT pathway related major genes were cloned successfully in silkworm. The analysis of conserved structural domains suggests that BmHOP is one of the JAK kinases, which phosphorylates the BmSTAT transcription factor and leads to activation of target genes. BmSOCS members, Bmken, BmPIAS 1 and BmPIAS2 are negative feedback regulators in the JAKJSTAT pathway.

关 键 词：家蚕 JAK/STAT通路 基因克隆 序列分析 

分 类 号：Q78[生物学—分子生物学]