家蚕orcokinin基因的选择性剪接与表达分析  

Alternative Splicing and Expression Analysis of orcokinin Gene in the Silkworm,Bombyx mori

作　　者：王平阳 赵巧玲 裘智勇[1,2] 沈兴家 夏定国[1,2] Wang Pingyang;Zhao Qiaoling;Qiu Zhiyong;Shen Xingjia;Xia Dingguo(Jiangsu Key Laboratory of Sericultrual Biology and Biotechnology,College of Biotechnology,Jiangsu University of Science and Technology,Zhenjiang Jiangsu 212018,China;Key Laboratory of Silkworm and Mulberry Genetic Improvement,Ministry of Agriculture and Rural Affairs,Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang Jiang-su 212018,China)

机构地区：[1]江苏科技大学生物技术学院,江苏省蚕桑生物学与生物技术重点实验室,江苏镇江212018 [2]中国农业科学院蚕业研究所,农业农村部桑蚕遗传改良重点实验室,江苏镇江212018

出　　处：《蚕业科学》2018年第6期849-858,共10页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.31372378);江苏省研究生创新计划项目(No.KYCX17-1857);江苏科技大学研究生创新计划项目(No.YCX16B-02);农业农村部蚕桑遗传改良重点实验室开放课题(No.KL201704);江苏省自然科学基金项目(No.BK20151322);江苏省高校自然科学研究重大项目(No.15KJA180001)

摘　　要：Orcokinin(OK)是节肢动物门特有的一种神经肽,该神经肽自从在小龙虾中发现以来,便陆续在节肢动物门其他物种中鉴定到,然而由于其结构的复杂性和功能的多样性,对OK基因的结构与作用机制知之甚少。利用RACE克隆、qRT-PCR等方法对家蚕OK基因(BmOK)的结构与表达进行分析。结果表明BmOK存在选择性剪接,形成2个转录本BmOKA和BmOKB;BmOKA由外显子exon1、exon2、exon3、exon5、exon6、exon7和exon8组成,全长BmOKB由外显子exon1、exon2、exon3和exon4组成。BmOKA由于选择性剪接又有4个亚型,分别为BmOKA1(CDS长度为534 bp)、BmOKA2(CDS长度为564 bp)、BmOKA3(CDS长度为570 bp)和BmOKA4(CDS长度为600 bp),占比为22∶13∶5∶1,4个亚型有相同的5′ UTR(62 bp)和3′ UTR(188 bp);BmOKB只有一种CDS(864 bp),5′ UTR序列与BmOKA相同(62 bp),但3′ UTR(168 bp)不同。BmOKA主要在家蚕腹神经索表达,在4龄眠后18 h表达量最低;BmOKB在头部和中肠的表达量相对较高,眠期表达量高于其他时期。BmOKA与斜纹夜蛾Orcokinin同源性最高,完整蛋白质前体的同源性为33.80%。本研究清晰地展示了家蚕BmOK基因的结构与表达模式,为后续分析BmOK基因的功能奠定了理论基础并提供实验依据。Orcokinin( OK) is a unique neuropeptide of Arthropoda. It has been identified subsequently from many other Arthropoda species after its first finding in Orconectes limosus. However,due to its complexity of structure and diversity of functions,it is rarely known about the structure and expression of OK. In this study,RACE cloning and qRTPCR methods were used for the structure and expression analysis of Bombyx mori OK( BmOK) gene. The results indicated that BmOK gene was alternatively spliced into two transcripts BmOKA and BmOKB. BmOKA consists of exon1,exon2,exon3,exon5,exon6,exon7 and exon8,and BmOKB consists of exon1,exon2,exon3 and exon4.Furthermore,BmOKA contains four subtype transcripts BmOKA1,BmOKA2,BmOKA3 and BmOKA4,their CDS length are 534 bp,564 bp,570 bp and 600 bp respec-tively,and the distribution ratio is 22 ∶ 13 ∶ 5 ∶ 1. The four subtypes have the same 62 bp 5’ UTR and 188 bp 3’ UTR. There is only one type of BmOKB,with CDS of 864 bp. It has the same 62 bp 5’ UTR with BmOKA but different( 168 bp) 3’UTR. The tissue expression profile showed that BmOKA was mainly expressed in ventral nerve cord. The lowest expression level of BmOKA was at 18 hours after forth molting. The expression of BmOKB was relatively higher in head and midgut than in other tissues,and higher in molting period than in other periods. BmOKA has the highest identity with OK from Spodoptera litura. Their complete protein precursors are 33. 80% identical. The above analysis shows clearly the structure and expression pattern of BmOK gene in silkworm,which provides a theoretical basis and experimental reference for subsequent analysis of the function of BmOK gene.

关 键 词：BmOK基因 选择性剪接 神经肽 家蚕 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q786[农业科学—畜牧兽医]