| 潘畅，张宇宏，谢佳沁，李浩森，庞虹*,2016,孟氏隐唇瓢虫气味结合蛋白ComOBP1基因的克隆和时空表达[J].环境昆虫学报,(2):249-260 |
| 孟氏隐唇瓢虫气味结合蛋白ComOBP1基因的克隆和时空表达 |
| Cloning and spatio-temporal expression of the odorant binding protein ComOBP1 gene from Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) |
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| DOI： |
| 中文关键词: 孟氏隐唇瓢虫 气味结合蛋白 CmonOBP1基因 基因克隆 序列分析 时空表达 |
| 英文关键词:Cryptolaemus montrouzieri Mulsant odorant binding protein CmonOBP1 gene gene cloning sequence analysis spatio-temporal expression |
| 基金项目:国家重点基础研究发展计划（973计划）项目（2013CB127600）；国家自然科学基金面上项目（31171899，31572052）；国家自然科学基金项目（31301720） |
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| 中文摘要: |
| 采用RT-PCR和RACE技术，从孟氏隐唇瓢虫 Cryptolaemus montrouzieri Mulsant中成功克隆出气味结合蛋白（ComOBP1）基因的全序列（Genbank 登陆号：KU170686）。ComOBP1基因全长922 bp，包括5′端长为40 bp的非编码区域（UTR），及3′端长为462 bp的UTR，开放阅读框ORF长为420 bp，编码139个氨基酸，预测的分子量为15.516 kDa，等电点pI为6.57，存在AATAAA加尾信号。N-末端疏水区包含由20个氨基酸构成的信号肽，无跨膜结构，有4个保守的半胱氨酸，属于Minus-C OBP，也是在孟氏隐唇瓢虫中发现的第一个Minus-C OBP。氨基酸序列中有且仅有一个N-糖基化位点为62 NLSA，并存在2个潜在的磷酸化位点。与其它昆虫的Minus-C OBPs进行同源性比较并构建系统发育树，发现与同为鞘翅目Coleoptera昆虫的同源性较高。利用Real-time PCR、RT-PCR技术对CmonOBP1基因在孟氏隐唇瓢虫不同发育阶段，不同组织，不同营养条件及不同食性下的表达水平进行了测定，结果显示，CmonOBP1基因在整个发育阶段均有表达，雄性成虫期具有最高表达量，且多在成虫的头部及翅部表达。当营养条件发生变化时，表达丰度不会发生变化，当猎物由天然猎物柑橘粉蚧 Planococcus citri Risso变成碗豆修尾蚜 Megoura japonica Matsumura时，表达量会明显下降。该结果表明，孟氏隐唇瓢虫的不同发育阶段、不同组织及猎物种类会影响CmonOBP1基因的表达，从而进一步影响其嗅觉行为。同时，CmonOBP1基因可能在雄虫相关的信息素感受过程中发挥着重要作用。 |
| 英文摘要: |
| The odorant binding protein gene CmonOBP1 (Genbank number: KU170686) from Cryptolaemus montrouzieri ouzieri Mulsant was cloned by technology of RT-PCR and RACE. The full length was 922 bp, containing a 5′ UTR (untranslated region) of 40 bp and a 3′UTR of 462 bp. ORF (Open Reading Frame) was 420 bp which encoded a polypeptide of 139 amino acids with an estimated molecular weight of 15.516 kDa and pI of 6.57. Besides, the sequence had an AATAAA tailing signal. The predicted N-terminal hydrophobic region containing 20 amino residues displayed the characteristic features of a signal peptide, and no transmembrane domain was predicted. Four conservative cysteine residues classified CmonOBP1 as the first Minus-C OBP which was found in C.montrouzieri. The amino acid sequence had only one N.glycosylation site 62 NLSA, and two potential phosphorylation sites. Compared with other insect Minus-C OBPs, CmonOPB1 was much identitier to the other Coleoptera. By Real-time PCR and RT-PCR methods, we determined the expression pattern of CmonOBP1 in different developmental stages, various adult tissues, different larval food regimes and other food. Spatio-temporal expression result showed that CmonOBP1 expressed in every stage and the male adult had the highest level. In addition, CmonOPB1 had a higher expression level in adult head and wing than that of other adult tissues. When larval diet changed either poor or rich, the expression level wouldn`t be influenced markedly, while when the food changed from natural prey. Planococcus citri Risso to Megoura japonica Matsumura, the expression level decreased significantly. The result suggested that the expression level of CmonOBP1 gene was development, tissues and prey type dependent, indicating a distinct sensitivity in olfactory. In addition, CmonOBP1 gene might play an important role in the process of male adult feeling related pheromone. |
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