意大利蜜蜂AmMETTL3基因的分子克隆与表达模式OACSTPCD

To clone the AmMETTL3 gene in Apis mellifera ligustica,analyze the physicochemical property and molecular characteristics of AmMETTL3 protein and determine the expression pattern of AmMETTL3 in different tissues and developmental stages of worker bees.AmMETTL3 CDS was amplified by PCR and then verified using Sanger sequencing.Related bioinformatic software were used to predict physicochemical property and molecular characteristics,identify structural domains and conserved motifs and construct phylogenetic tree of AmMETTL3.RT-qPCR was employed to detect the relative expression levels of AmMETTL3 in seven different tissues and five various developmental stages.The results indicated that the CDS of AmMETTL3 was successfully cloned,AmMETTL3 protein do not have typical transmembrane domain and signal peptide,and METTL3 is highly conserved in various insects including one structure domain MT-A70 and three same conserved motifs.AmMETTL3 and A.cerana METTL3 were clustered into one group in the phylogenetic tree.The expression level of AmMETTL3 was significantly up-regulated in hypopharyngeal gland(P＜0.01),significantly down-regulated in midgut(P＜0.01)compared with that in antennae.The expression level of AmMETTL3 in 3-day-old larvae(P＜0.01)was significantly lower than that in eggs.The expression level of AmMETTL3 in 12-day-old pupa(P＜0.01)was significantly higher than that in eggs;the expression level of AmMETTL3 in workers at 2,6,12,15 and 18 day-old(P＜0.01)was significantly lower than that in workers at 1 day-old.This study provides references and a basis for further investigating the function of AmMETTL3 and itsrole in epigenetic regulatory mechanisms.