刊期:双月刊
主管单位:中国科学院
主办单位:中国科学院动物研究所,中国昆虫学会
地址:北京市朝阳区北辰西路1号院5号中国科学院动物研究所
邮编:100101
电话:010-64807137
传真:010-64807137
E-Mail:entom@ioz.ac.cn
刊号:ISSN 2095-1353
        CN 11-6020/Q
国内发行代号:2-151
国际发行代号:BM-407
发行范围:国内外公开发布
定价:138元/册
定价:828元/年
银行汇款:中国工商银行北京海淀西区支行
户名:中国科学院动物研究所
帐号:0200 0045 0908 8125 063

您所在位置:首页->过刊浏览->2021年58卷第3期



九香虫保幼激素环氧水解酶基因克隆、生物信息学及表达分析
Cloning, bioinformatics and expression of the juvenile hormone epoxide hydrolase gene in Aspongopus chinensis Dallas
周汶桢 吴有芳 郭建军
点击:696次 下载:30次
DOI:10.7679/j.issn.2095-1353.2021.062
作者单位:贵州大学昆虫研究所,贵州山地农业病虫害重点实验室,贵阳 550025
中文关键词:保幼激素环氧水解酶;九香虫;克隆;生物信息学分析;表达分析
英文关键词:juvenile hormone epoxide hydrolase; Aspongopus chinensis; gene clone; bioinformatics analysis; expression analysis
中文摘要:
【目的】 保幼激素环氧水解酶(Juvenile hormone epoxide hydrolase,JHEH)是昆虫体内保幼激素(Juvenile hormone,JH)的主要降解酶。本文旨在分析九香虫Aspongopus chinensis Dallas保幼激素环氧水解酶基因序列(AcJHEH)信息,探索其在九香虫生长发育过程中的作用。【方法】 以九香虫成虫的cDNA为模板,采用RT-PCR技术克隆获得AcJHEH基因序列,并利用生物信息学软件对其编码蛋白的理化特性、结构特征和系统进化进行分析;采用qRT-PCR技术检测并分析九香虫不同龄期和不同组织中AcJHEH的相对表达量。【结果】 成功获得了AcJHEH的完整开放阅读框ORF序列,全长均为1 363 bp,共编码453个氨基酸,预测分子量为51.74 ku,等电点(pI)为7.66,分子蛋白式C2414H3683N581O653S14,含有N-端跨膜基序XWG、催化三联体、保守基序HGWP和2个酪氨酸,属于环氧水解酶家族。系统进化树分析表明,AcJHEH与茶翅蝽Halyomorpha halysJHEH聚为一支,再与同为半翅目的臭虫Cimex lectulariu和绿盲蝽Apolygus lucorumJHEH聚为一类。荧光定量PCR结果显示,AcJHEH在九香虫不同发育阶段和各组织中均有表达,且脂肪中的表达量极显著高于其它组织(P<0.001);滞育九香虫成虫表达量最高,其次为4-5龄若虫都极显著高于其它发育阶段(P<0.001)。【结论】 九香虫JHEH属于环氧水解酶家族,确定为保幼激素环氧水解酶。依据qPCR结果推测AcJHEH通过改变九香虫体内保幼激素浓度来影响九香虫4-5龄若虫蜕皮发育、成虫生殖系统发育和滞育等。
英文摘要:
[Objectives]  To sequence the Aspongopus chinensis juvenile hormone epoxide hydrolase and use bioinformatic analysis to investigate its role in the development of A. chinensis. [Methods]  The cDNA sequence of AcJHEH was amplified and cloned from adult A. chinensis with RT-PCR. Its physiochemical properties and structural characteristics were described and a phylogenetic tree of the deduced amino acids constructed using bioinformatics software. The expression of AcJHEH in different tissues (ovary, testis, fat, muscle, body wall, midgut and head) and in different developmental stages (1st, 2nd, 3rd, 4th, 5th instar larvae, adult males and females, diapausing and reproductive adults) was measured using quantitative real-time PCR. [Results]  The entire ORF sequence of AcJHEH containing 1 363 bp was obtained. Both genes encoded a putative protein of 453 amino acids with an estimated molecular weight of 51.74 ku. The isoelectric point (pI) was 7.66 and the molecular protein formula was C2414H3683N581O653S14. The amino acid encoded by JHEH contained the N-terminal transmembrane motif XWG, a catalytic triad, the conserved motif HGWP and two tyrosines, suggesting that it belongs to the epoxide hydrolase family. The phylogenetic tree indicates that AcJHEH is most similar to Halyomorpha halys JHEH and that it clusters with JHEH genes from the Hemipteran species Cimex lectulariu and Apolygus lucorum. Quantitative real-time PCR revealed that AcJHEH is expressed in various tissues and developmental stages of A. chinensis. Expression in the fat body was significantly higher than in other tissues (P<0.001). AcJHEH was most highly expressed in diapausing A. chinensis, followed by 4-5 instar larvae, in which expression was significantly higher than in other instars (P<0.001). [Conclusion]  The JHEH of A. chinensis belongs to the epoxide hydrolase enzyme family and was identified as a juvenile hormone epoxide hydrolase. qPCR results suggest it is involved in the molting and development of 4-5th instar larvae, the development of the adult reproductive system and diapause.
读者评论

      读者ID: 密码:   
我要评论:
版权所有©2024应用昆虫学报》编辑部 京ICP备10006425号
本系统由北京菲斯特诺科技有限公司设计开发
您是本站第9020977名访问者