西伯利亚蝗酚氧化酶原2(PPO2)基因的克隆及表达分析  

Cloning and Expression Analysis of Prophenoloxidase(PPO2)Gene of the Gomphocerus sibiricus

作　　者：阿地拉·阿吉 廖杰杰 马爱霞 魏笑佳 叶小芳 季荣 扈鸿霞 ADILA A-Ji;LIAO Jie-Jie;MA Ai-Xia;WEI Xiao-Jia;YE Xiao-Fang;JI Rong;HU Hong-Xia(Key Laboratory of Special Environment Biodiversity Application and Regulation in Xinjiang/International Center for the Collaborative Management of Cross-border Pest in Central Asia/College of Life Sciences,Xinjiang Normal University,Urumqi 830054,China)

机构地区：[1]新疆师范大学,生命科学学院/中亚区域跨境有害生物联合控制国际研究中心/新疆特殊环境物种多样性应用与调控实验室,乌鲁木齐830054

出　　处：《农业生物技术学报》2021年第11期2189-2197,共9页Journal of Agricultural Biotechnology

基　　金：新疆维吾尔自治区高校科研计划项目(XJEDU2021I022);国家级大学生创新训练项目(202010762014);新疆维吾尔自治区天山英才计划;新疆师范大学校级招标课题(XJTSWZ-2021-03)。

摘　　要：虫酚氧化酶(phenoloxidase,PO)是一类重要的免疫效应分子,属于3型铜蛋白家族,具有氧化还原酶活性。PO通常以其非活化状态的前体酚氧化酶原(prophenoloxidase,PPO)的形式存在于昆虫血淋巴中,经蛋白酶水解激活催化黑色素合成和酚类氧化成醌。西伯利亚蝗(Gomphocerus sibiricus)是新疆高山、亚高山草原地区的优势种。本研究采用cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术克隆了西伯利亚蝗PPO2基因的全长cDNA序列。该序列全长2439 bp(GenBank No.KY981766);生物信息学分析结果显示PPO2基因与东亚飞蝗(Locusta migratoria)和红褐斑腿蝗(Diabolocatantops pinguis)的酚氧化酶原的同源性最高;有两个铜绑定位点及保守的组蛋白残基,具有保守的酪氨酸酶结构域和蛋白水解酶的作用位点。PPO2基因表达分析结果表明,该基因主要在成虫后肠表达,其次为中肠,在蝗虫马氏管和唾液腺中的表达没有差异,在脂肪体中表达最少。本结果为研究PPO在蝗虫体内的组织分布及免疫防疫功能提供了基础数据。Phenoloxidases(POs)are important immune effector molecules of insect and belong to the type 3 copper protein familywith oxidoreductase activities.Generally,PO is present as a zymogen,prophenoloxidase(PPO),which is activated by a serine protease cascade upon recognition of foreign invaders,and then catalyzes both melanin biosynthesis and the oxidation of phenols to quinones.Gomphocerus sibiricus is dominant pest in alpine and subalpine grassland areas in Xinjiang.The study of immune related gene of G.sibiricus can lay a foundation for the future study of its interaction with pathogenic organisms to carry out biological control.The full length cDNA of PPO2 were cloned by RACE technology in this study,and which consisted of 2439 bp(GenBank No.KY981766).Bioinformatic analysis indicated that PPO2 is highly similar to Locusta migratoria and Diabolocatantops pinguis.It contained 2 common copper-binding regions,2 potential proteolytic activation sites,a plausible thiolester site,and a conserved C-terminal region.Genetic expression analysis showed that the gene expression level was the highest inhindgut of G.sibiricus.The expression level in posterior intestine was significantly higher than midgut,and there were no difference between salivary glands and malpighian tubules,the expression was least in fat body.This study could provide a theoretical basis for further study of the function of PPO2 gene in G.sibiricus.

关 键 词：西伯利亚蝗 酚氧化酶原2(PPO2)基因 序列分析 表达分析 

分 类 号：S433[农业科学—农业昆虫与害虫防治]