Sequence and Expression Analysis of ADP-ribosylation Factor 1 Gene from Monochamus alternatus

CHEN Jingxiang; CHENG Jie; LIN Tong

doi:10.15933/j.cnki.1004-3268.2017.07.010

Abstract：To explore the expression characteristics of ADP-ribosylation factor 1 gene (ARF1)from Ras supergene family in the process of insect development and under temperature stress,ARF1 gene was filtrated from cDNA library of Monochamus alternatus,as the research object,and was named MaARF1 (GenBank:KY368168),whose sequence and expressional characteristic in each insect state or under different temperature stress were analyzed in this study.The results showed that the coding sequence of MaARF1 was 549 bp in length,encoding 182 amino acids.The predicted secondary structure of MaARF1 was mainly composed of alpha helix and beta sheet,followed by random curl and beta turn.Amino acids sequence multiple alignment analysis of MaARF1 showed the highest homology with Tribolium castaneum (100％) and there were one myristoylation (G2) and five conservative domains(G1 box-G5 box).By RTqPCR technology,gene expression analysis showed that MaARF1 expression level was associated with holometabolous development of M.alternatus and it was expressed in each insect state,continuously.The expression level was relatively higher in second and fifth instar larvae,showed a rising trend in larvalpupal stage,a downward trend during eclosion,and reached the maximum amount in the following day of pupa.MaARF1 under different temperature stress was expressed,low temperature and high temperature would lead to lower expression level,when the temperature was higher or lower,falling of expression level was more significant.Therefore,MaARF1 may mainly affect two key metamorphosis point,nymphosis and eclosion,so as to affect the entire development process in M.alternatus,and it may also be involved in the Ca2+ signaling pathways depending on the temperature.