野桑蚕小分子热激蛋白基因BmmHSP19.9的克隆与表达分析  

Cloning and Expression of Small Heat Shock Protein Gene Bmm-HSP19.9 in Bombyx mandarina (Lepidoptera: Bombycidae)

作　　者：丁佳奇 蒋青萍 陈恩熙 林明君 钱荷英 赵国栋[1,2] Ding Jiaqi;Jiang Qingping;Chen Enxi;Lin Mingjun;Qian Heying;Zhao Guodong(School of Biotechnology,Jiangsu University of Science and Technology,Zhengjiang Jiangsu 212100,China;Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhengjiang Jiangsu 212100,China)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212100 [2]中国农业科学院蚕业研究所,江苏镇江212100

出　　处：《蚕业科学》2023年第6期481-489,共9页ACTA SERICOLOGICA SINICA

基　　金：财政部和农业农村部国家现代农业产业技术体系项目;江苏省自然科学基金面上项目(BK20201229)。

摘　　要：为探究野桑蚕(Bombyx mandarina)响应外源不良胁迫的分子机制,利用RT-PCR技术克隆野桑蚕小分子热激蛋白(small heat shock protein, sHSP)基因BmmHSP19.9,并进行生物信息学分析。BmmHSP19.9基因(GenBank登录号:OP779245)的开放阅读框(ORF)长534 bp,编码177个氨基酸。BmmHSP19.9具有sHSP特征结构域,相对分子质量为19.9 kD,等电点(pI)为6.53。同源比对和系统进化分析显示,昆虫sHSP蛋白具有较高的保守性,BmmHSP19.9与家蚕的BmHSP19.9亲缘关系最近。RT-qPCR结果表明,BmmHSP19.9在野桑蚕5龄幼虫的精巢、中部丝腺和后部丝腺的表达量较高;在高温(40℃)和低温(8℃)胁迫时,BmmHSP19.9在5龄幼虫所有组织中的表达水平均有所上调;添食BmNPV对BmmHSP19.9在5龄幼虫血淋巴和中肠组织的表达有诱导作用;溴氰菊酯胁迫下,BmmHSP19.9在5龄幼虫中肠中的表达水平显著上调。上述结果表明,BmmHSP19.9可能在野桑蚕响应外源不良胁迫的过程中发挥了重要作用。In order to explore the molecular mechanism of Bombyx mandarina in response to exogenous adverse stresses, the small heat shock protein(sHSP) gene BmmHSP19.9(GenBank accession No. : OP779245) of B. mandarina was cloned by RT-PCR, and analyzed by bioinformatic methods. Its open reading frame(ORF) is 534 bp in length, encoding 177 amino acids. BmmHSP19.9 has the sHSP characteristic domain. The relative molecular weight was 19.9 kD and the isoelectric point(pI) is 6.53. Homologous comparison and phylogenetic analysis showed that the insect sHSP proteins are highly conservative. The relationship between BmmHSP19.9 and BmHSP19.9 from Bombyx mori was the closest. RT-qPCR result showed that BmmHSP19.9 was highly expressed in testis, middle and posterior silk glands of the 5th instar larvae. The expression level of BmmHSP19.9 was increased in all tissues under high(40 ℃) and low(8 ℃) temperature stresses. BmNPV feeding could induce the expression of BmmHSP19.9 in hemolymph and midgut. The expression level of Bmm-HSP19.9 was significantly up-regulated in the midgut of the 5th instar larvae by feeding with deltamethrin. The results showed that BmmHSP19.9 may play an important role in the response to exogenous adverse stress in B.mandarina.

关 键 词：野桑蚕 小分子热激蛋白基因 基因克隆 表达分析 外源胁迫 

分 类 号：Q966[生物学—昆虫学] Q786