枣实蝇气味结合蛋白基因的克隆及表达谱分析  

Cloning and Expression Profiling of Odorant Binding Protein Gene in Carpomya vesuviana(Costa)(Diptera:Tephritidae)

作　　者：李亚伟[1,2,3] 张小菊[2] 王科珂 于艳雪 张祥林[2] 张伟[2] 张俊华 杨定[3] 陈乃中 LI Ya - wei ZHANG Xiao - ju WANG Ke - ke YU Yan - xuc ZHANG Xiang- lin ZHANG Wei ZHANG Jun- hua YANG Ding CHEN Nai -zhong(Research Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China Xinjiang Entry Exit Inspection and Quarantine Portal, Urumqi 830063, China China Agricultural University, Beijingl O0083 , China)

机构地区：[1]中国检验栓疫科学研究院植物检疫研究所,北京100176 [2]新疆出入境检验检疫局,乌鲁木齐830063 [3]中国农业大学,北京100083

出　　处：《新疆农业科学》2017年第2期313-319,共7页Xinjiang Agricultural Sciences

基　　金：国家质检总局科研项目“重要检疫性入侵害虫枣实蝇监测诱集物质”(2015IK321);国家“十二五”科技支撑计划“重要外来有害生物传入风险及监测技术”(2015BAD08B01);质检公益性行业科研专项“外来生物定殖因子及危害因子研究及防控技术”(201410014)~~

摘　　要：【目的】克隆枣实蝇Carpomya vesuviana Costa气味结合蛋白(odorant binding protein,Cvco OBP)基因,对其表达谱进行分析,为枣实蝇气味结合蛋白的深入研究奠定基础。【方法】对枣实蝇头部RNA进行转录组测序,通过蛋白序列比对以及构建系统进化树分析枣实蝇OBP序列,运用半定量反转录-聚合酶链反应(semi-quantitative reverse transcription and polymerase chain reaction,sqRT-PCR)方法对获得的Cvco OBP序列进行组织表达特异性研究。【结果】转录组测序后共获得7个OBP,命名为(Gen Bank登录号分别为:KU975053、KU975054、KU975055、KX059394、KU975056、KX059395、KX059396)。对Cvco OBP1-7进行蛋白序列比对发现,Cvco OBP1、Cvco OBP3、Cvco OBP4、Cvco OBP6、Cvco OBP7是典型的OBP家族成员,具有6个高度保守的半胱氨酸残基;Cvco OBP2和Cvco OBP5属于"Minus-C"家族成员,具有4个高度保守的半胱氨酸残基。系统进化分析结果显示,Cvco OBP1-7分别与不同的OBP有较近的进化关系。半定量PCR检测结果发现,Cvco OBP1-7在枣实蝇腹部、头部及胸部都有表达,且表达量较高;Cvco OBP1、Cvco OBP2在翅中有表达,但表达量相对较低;Cvco OBP1、Cvco OBP5、Cvco OBP6在足中也有表达,但表达量相对较低。【结论】获得并分析了7个Cvco OBP,不同Cvco OBP表达部位和表达量各不相同;Cvco OBP1-6分别属于OBP56g,OBP8a,OBP83a,OBP56h,OBP99c,OBP83ef类型。[ Objective] This study aims to clone Carpomya vesuviana Costa odorant binding protein （Cv- eoOBP） and analyze expression profile of CveoOBP in order to lay the foundation of researching CveoOBP in Carpomya vesuviana Costa. [ Method ] Based on transeriptome sequencing of cephalic RNA, the study has analyzed CveoOBP according to alignment and phylogenetie analysis. The research was also conducted into the tissue specific expression of CveoOBP by semi - quantitative reverse transcription and polymerase chain reaction （sqRT- PCR）. [ Result] The study has obtained seven CveoOBP that had been named respectively as CvcoOBP1- 7 （GenBank accession number： KU975053, KU975054, KU975055, KX059394, KU975056, KX059395 and KX059396）. According to amino acid alignment, CvcoOBP1, CvcoOBP3, CveoOBP4, CvcoOBP6 and CvcoOBP7 had been proved to be a family of classic OBP, which contained six highly conversed eysteine residues. CveoOBP2 and CveoOBP5 belonged to a family of ＂Minus - C＂ , which had only four highly conversed eysteine residues. Phylogenetie analysis showed that CveoOBP1 -7 had close relationships with different OBPs. Finally, sqRT- PCR revealed that CveoOBP1 -7 could be highly expressed in abdomen, head and thorax. CvcoOBP1 and CvcoOBP2 could be also expressed in wings with relatively low expression. CvcoOBP1, CvcoOBP5 and CvcoOBP6 were also found in legs. [ Conclusion ] For the first time, the study obtained and analyzed seven CveoOBP. Different CveoOBP are various in terms of expression sites and expression quantity. The CveoOBP1 -6 separately belongs to OBP56g, OBP8a, OBP83a, OBP56h, OBP99e and OBP83ef type.

关 键 词：枣实蝇 气味结合蛋白 组织表达 

分 类 号：S436.65[农业科学—农业昆虫与害虫防治] S188[农业科学—植物保护]