中国农业大学学报2024，Vol.29Issue(1)：99-108,10.DOI:10.11841/j.issn.1007-4333.2024.01.09
意大利蜜蜂AmMETTL3基因的分子克隆与表达模式
Molecular cloning and expression pattern determination of AmMETTL3 gene in Apis mellifera ligustica
摘要
Abstract
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.关键词
意大利蜜蜂/N6-甲基腺苷/甲基化转移酶3/MT-A70/分子特性/表达模式Key words
Apis mellifera ligustica/N6-methyladenosine/methyltransferase 3/MT-A70/molecular characteristics/expression pattern分类
农业科技引用本文复制引用
刘小玉,付中民,郭睿,刘治滩,荆欣,冯佩林,高旭泽,吴鹰,李琪明,樊念,陈大福..意大利蜜蜂AmMETTL3基因的分子克隆与表达模式[J].中国农业大学学报,2024,29(1):99-108,10.基金项目
国家自然科学基金项目(32172792,31702190) （32172792,31702190）
国家现代农业产业技术体系专项资金项目(CARS-44-KXJ7) （CARS-44-KXJ7）
福建省自然科学基金面上项目(2022J01131334) （2022J01131334）
福建农林大学科技创新专项基金项目(KFb22060XA) （KFb22060XA）
福建农林大学硕士生导师团队项目 （）
福建农林大学动物科学学院(蜂学学院)科研扶持项目 （蜂学学院）
