草地贪夜蛾钙粘蛋白重复区片段的克隆表达及时空分析  

Cloning,expression and temporal analysis of Cadherin in Spodoptera frugiperda(Lepidoptera:Noctuidae)

作　　者：王靖怡[1] 李颖[1] 顾俊文 张珀瑞 祖尔东·加拉力丁 张琪[1] WANG Jing-Yi;LI Ying;GU Jun-Wen;ZHANG Po-Rui;ZUERDONG·Jalaliding;ZHANG Qi(College of Plant Protection,Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[1]沈阳农业大学植物保护学院,沈阳110866

出　　处：《环境昆虫学报》2024年第5期1171-1180,共10页Journal of Environmental Entomology

基　　金：国家自然科学基金(32072482);辽宁省教育厅青年基金(LSNQN202017)。

摘　　要：为探明钙粘蛋白在草地贪夜蛾Spodoptera frugiperda中不同发育阶段、不同组织中的表达水平,进一步探索草地贪夜蛾钙粘蛋白作为Cry毒蛋白受体的功能性作用,本研究克隆并原核表达了草地贪夜蛾钙粘蛋白(Cadherin)SfCR10-12片段,以及SfCR10,SfCR11,SfCR123个分段重复区片段,并对其时空表达模式进行了分析。结果表明:(1)草地贪夜蛾与甜菜夜蛾Spodoptera exigua钙粘蛋白序列高度相似达80.46%。(2)获得的SfCR10-12序列(GenBank登录号:OP547784)长度为933 bp,开放阅读框810 bp,编码268个氨基酸。(3)SfCR10-12基因在不同发育阶段均有表达,在5龄和6龄幼虫中表达量最高,此外在草地贪夜蛾中肠部位有一定的表达,但是在头部和体壁表达量较低。(4)经原核表达和纯化后获得了分子大小约为34 kDa、9.57 kDa、14.11 kDa和11.06 kDa的SfCR10-12、SfCR10、SfCR11和SfCR12蛋白片段。本研究结果为深入研究草地贪夜蛾钙粘蛋白不同重复区介导Bt Cry毒素对草地贪夜蛾的作用机制提供了基础。In order to investigate the relative transcript level of cadherin gene in Spodoptera frugiperda in different tissues and developmental stages,and further investigate the functional role of cadherin as Cry toxin receptor,we cloned and over-expressed cadherin fragment 10-12 of S.frugiperda(SfCR10-12)along with three truncated fragments SfCR10,SfCR11,SfCR12,and analyzed its temporal and spatial expression levels in S.frugiperda.Results showed that:(1)The cadherin sequence of S.frugiperda presents highest homology with that in S.exigua(80.46%).(2)The SfCR10-12 gene was 933 bp(GenBank accession number OP547784),with ORF of 810 bp encoding 268 amino acids.(3)The SfCR10-12 gene was detected to express in different developmental stages with significantly higher expression in 5th and 6th instar larvae.(4)The molecular size of SfCR10-12,SfCR10,SfCR11 and SfCR12 protein fragments were approximately 34 kDa,9.57 kDa,14.11 kDa and 11.06 kDa,respectively.Our results provide fundamental knowledge for the essential role of cadherin as a functional receptor of Cry toxin underlying its mode of action in S.frugiperda.

关 键 词：草地贪夜蛾 钙粘蛋白 克隆 蛋白表达 时空表达 

分 类 号：Q968.1[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]