中华蜜蜂磷酸丙糖异构酶基因克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of Gene Encoding Triose Phosphate Isomerase of Apis cerana cerana

作　　者：樊兆斌 张淑萍 葛中东 周宛蓉 张凯 夏宇欣 姜莉莉 蒋培红 FAN Zhao-bin;ZHANG Shu-ping;GE Zhong-dong;ZHOU Wan-rong;ZHANG Kai;XIA Yu-xin;JIANG Li-li;JIANG Pei-hong(College of Pharmacy,Heze University,Heze 274015,China;Heze Institute of Food and Drug Inspection,Heze 274000,China)

机构地区：[1]菏泽学院药学院,山东菏泽274015 [2]菏泽市食品药品检验检测研究院,山东菏泽274000

出　　处：《中国兽医杂志》2020年第11期17-21,F0002,共6页Chinese Journal of Veterinary Medicine

基　　金：山东省自然科学基金项目(ZR2017MC055);山东省高等学校科研计划项目(J17KA126);国家大学生创新创业训练计划项目(201710455149);菏泽学院博士基金(XY16BS08)。

摘　　要：本试验克隆中华蜜蜂磷酸丙糖异构酶(Tpi)基因,并对其编码产物进行生物信息学分析预测。利用RT-PCR方法扩增中华蜜蜂幼虫Tpi基因(AcTpi),并对其核苷酸和推导氨基酸序列进行遗传演化分析,利用生物信息学软件对AcTpi编码的蛋白功能进行分析和预测。预测分析结果表明,AcTpi具有完整的开放阅读框(ORF),CDS区为744 bp,编码247个氨基酸。理论分子量为26.88 kDa,理论等电点为7.76,不稳定系数为27.32,脂肪系数为93.52,平均亲水指数为-0.182。存在3个明显的疏水区,无信号肽,不存在跨膜区;其编码蛋白较稳定,为亲水性蛋白。二级结构预测显示,AcTpi编码的蛋白以无规则卷曲为主,存在多个α螺旋和β折叠区域。本试验成功克隆了中华蜜蜂幼虫Tpi基因,为深入研究AcTpi功能提供依据,为进一步阐明AcTpi在糖酵解过程中的作用及其功能的开发应用提供了科学依据。In order to clone the triose phosphate isomerase(Tpi)gene of Apis cerana cerana and analyze its physicochemical property and secondary structure.RT-PCR was used to amplify AcTpi gene.The genetic evolution of nucleotides and derived amino acid sequences were analyzed,and bioinformatics softwares were used to predict and analyze the structure and function of AcTpi coden protein.The results of bioinformatics analysis showed that the AcTpi gene included a 744 bp whole length CDS(coding 247 amino acids).The relative molecular weight,isoelectric point,instability index and fat coefficient of AcTpi were 26.88 kD,7.76,27.32 and 93.52,respectively.The prediction of AcTpi structure showed that AcTpi had neither signal peptide,transmembrane domain nor transmembrane domain,while it had 3 obvious hydrophobic domains,and the average hydrophilic index was-0.182.The prediction of secondary structure revealed that someα-helixes andβ-sheets existed in AcTpi protein while random coil was the major pattern.This study indicates that the whole CDS sequence of AcTpi gene is successfully cloned,and the bioinformatics analysis can be used as a reference for the further study on the function of AcTpi.This study provides an important foundation for further elucidating the role of AcTpi in glycolysis and its application in other functions.

关 键 词：中华蜜蜂 磷酸丙糖异构酶基因 基因扩增 生物信息学 

分 类 号：S896.1[农业科学—特种经济动物饲养]