中、意蜂Atg12基因克隆表达及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2019.03.001

Abstract

Abstract:

The purpose of this study was to investigate the expression differences of autophagy-related 12 (Atg12) gene in Apis cerana cerana and Apis mellifera,and provide references for studying the mechanism of Apis cerana cerana resitance to Varroa destructor.The primers were designed according to the Atg12 gene sequence of Apis cerana in GenBank (accession No.:XM_017048509.1).The Atg12 gene was amplified from the head of Apis cerana cerana and Apis mellifera,then was cloned,sequenced and expressed prokaryoticly,and the amino acid sequence and protein structure were analyzed.Meanwhile,Real-time PCR was used to analyze the different expression of Atg12 gene in head tissues of Apis cerana cerana and Apis mellifera.The results showed that the target fragment size was 450 bp and the recombinant protein size was about 42 ku.Genetic phylogenetic tree analysis showed that the relationship among Apis cerana cerana,Apis mellifera,Apis dorsata and Apis florea was closer in the evolution of Atg12 gene.Bioinformatics analysis of the recombinant protein showed that the secondary structure of the protein contained 6 polypeptide binding sites,6 β-sheets and 4 α-helices,the molecular weight was about 16.13 ku,and the isoelectric point was 6.73.The results of Real-time PCR showed that the expression of Atg12 gene in the head tissue of Apis cerana cerana was extremely significantly higher than that of Apis mellifera.The results of this study provided references for further study on the mechanism of action of Atg12 gene in Apis cerana cerana resitance to Varroa destructor.

Key words: Apis cerana cerana; Apis mellifera; Atg12 gene; cloning; prokaryotic expression; resistance to Varroa destructor

CLC Number:

TU Yangyang, WU Jiangli, WU Pengjie, TAN Jing, YU Huimin, XU Jin, GUO Yueqin, WANG Lihua, XU Shufa. Cloning,Expression and Bioinformatics Analysis of Atg12 Gene in Apis cerana cerana and Apis mellifera[J]. , 2019, 46(3): 643-651.

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Cloning,Expression and Bioinformatics Analysis of Atg12 Gene in Apis cerana cerana and Apis mellifera

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