中华蜜蜂气味结合蛋白基因AcerOBP7的克隆及序列分析

Cloning and sequence analysis of odorant-binding protein gene AcerOBP7 from Apis cerana cerana

[目的]克隆中华蜜蜂(Apis cerana cerana)AcerOBP7基因并分析其相关生物学信息。[方法]收集中华蜜蜂样本,提取总RNA,反转录制备cDNA。设计特异性引物,PCR扩增AcerOBP7基因的ORF序列。利用多种生物信息学软件对编码蛋白的理化性质和结构特征进行预测和分析。[结果]获得中华蜜蜂气味结合蛋白基因AcerOBP7的cDNA序列。AcerOBP7基因的开放阅读框(ORF)长438 bp,编码145个氨基酸。存在信号肽,无跨膜结构,在53~140个氨基酸之间有一个昆虫气味结合蛋白家族的PBP-GOBP家族的保守结构域,信号肽源于氨基酸中的1~20位,其对应的剪切位点落在氨基酸中的20位与21位间。有一些相对清晰的疏水区。[结论]AcerOBP7蛋白属于Classical OBP亚家族,具有典型的PBP-GOBP家族结构,是一种分泌蛋白。

[Objective]To clone the Apis cerana cerana AcerOBP7 gene,and analyze its related biological information.[Method]The Apis cerana cerana samples were collected to extract total RNA,and cDNA was prepared by reverse transcription.Specific primers were designed,and the ORF of AcerOBP7 gene was amplified by PCR.The physicochemical and structural features of the encoded protein were predicted and analyzed using a variety of bioinformatics software.[Result]The cDNA sequence of AcerOBP7 gene of Apis cerana cerana was cloned and obtained.The open reading frame(ORF)of AcerOBP7 is 438 bp long and encodes 145 amino acids.There is a signal peptide,no transmembrane structure,and there is an insect odorant binding protein family PBP-GOBP superfamily conserved domain between amino acids 53-140;the signal peptide is located at amino acids1-20,and the amino acid splicing site is between 20 and 21.There are a number of relatively obvious hydrophobic regions.[Conclusion]AcerOBP7 belongs to the Minus-C OBP subfamily and has a typical structure of the PBP-GOBP family.AcerOBP7 is characterized by a secreted protein.