枣实蝇气味结合蛋白基因的克隆及表达谱分析

发布时间：2018-07-24 20:48

【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。
[Abstract]:[objective] to clone the Carpomya vesuviana Costa odor binding protein (odorant binding) OBP gene of jujube fly and analyze its expression profile, so as to lay a foundation for the further study of the odour binding protein. [methods] RNA in the head of jujube fruit fly was sequenced. OBP sequences of jujube fly were analyzed by protein sequence alignment and phylogenetic tree construction. Semi-quantitative reverse transcription and polymerase chain reaction-polymerase chain reaction (RT-PCR) was used to study the tissue expression specificity of the obtained Cvco OBP sequence. [results] Seven OBPs were obtained after transcriptome sequencing, named as (Gen Bank accession number KU975053, KU975054, KX059394KU975056, KX059395KX05939596). Protein sequence alignment of Cvco OBP1-7 revealed that Cvco OBP3Cvco OBP4Cvco OBP6Cvco OBP7 is a typical member of OBP family, with six highly conserved cysteine residues, Cvco OBP2 and Cvco OBP5 belonging to the "Minus-C" family, with four highly conserved cysteine residues. Phylogenetic analysis showed that Cvco OBP1-7 had a close evolutionary relationship with different OBP. The results of semi-quantitative PCR showed that Cvco OBP1-7 was expressed in the abdomen, head and chest of jujube fly, and Cvco OBP1Cvco OBP2 was expressed in the wings of the jujube fly, but the expression of Cvco OBP1, Cvco OBP5Cvco OBP6 was also found in the foot. But the expression level was relatively low. [conclusion] Seven Cvco OBPs were obtained and analyzed. The different Cvco OBP expression sites and expression levels were different. The OBP1-6 belonged to the OBP56gOBP8aOBP83hmh OBP99cOBP83ef type.
【作者单位】：中国检验检疫科学研究院植物检疫研究所;新疆出入境检验检疫局;中国农业大学;
【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)~~
【分类号】：S433

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