春尺蠖保幼激素结合蛋白AcinJHBP基因的克隆、分子特征与表达分析  

Cloning,molecular characteristics and expression analysis of juvenile hormone binding protein gene AcinJHBP in Apocheima cinerarius

作　　者：朱圣杰 崔阔澍 白松林 张鹏飞 郝水源 郝雅娴 陈龙 ZHU Sheng-jie;CUI Kuo-shu;BAI Song-lin;ZHANG Peng-fei;HAO Shui-yuan;HAO Ya-xian;CHEN Long(Agriculture Department,Hetao College,Bayannur,Inner Mongolia 015000,China;Hetao Area Green Agricultural Product Safety Production and Early Warning Control Laboratory,Bayannur,Inner Mongolia 015000,China;Sichuan Agricultural Technology Extension Station,Chengdu 610041,China;Ordos Afforestation Station,Ordos,Inner Mongolia 010700,China;Erenhot Center For Disease Control and Prevention,Erenhot,Inner Mongolia 011100,China)

机构地区：[1]河套学院农学系,内蒙古巴彦淖尔015000 [2]河套地区绿色农产品安全生产与预警控制实验室,内蒙古巴彦淖尔015000 [3]四川省农业技术推广总站,成都610041 [4]鄂尔多斯市造林总场,内蒙古鄂尔多斯010700 [5]二连浩特市疾病预防控制中心,内蒙古二连浩特011100

出　　处：《西南农业学报》2024年第7期1538-1544,共7页Southwest China Journal of Agricultural Sciences

基　　金：内蒙古自然科学基金项目(2021BS03028);内蒙古自治区高等学校科学研究项目(NJZY21197);“科技兴蒙”行动重点专项(NMKJXM202201)。

摘　　要：[目的]探究AcinJHBP基因在春尺蠖生长发育与滞育中的作用。[方法]基于前期所测春尺蠖蛹转录组数据(非滞育蛹-滞育蛹与滞育蛹-滞育解除蛹),克隆获取JHBP基因,命名为AcinJHBP(GenBank登录号:OR860357);利用生物信息学软件分析AcinJHBP基因的序列特征及理化性质;采用qPCR技术检测AcinJHBP的时空表达(不同发育阶段和组织)情况。[结果]春尺蠖AcinJHBP基因完整CDS序列全长726 bp,编码241个氨基酸,蛋白质预测分子量为26.97 kDa,预测等电点为5.13。春尺蠖AcinJHBP基因预测分子式为C_(1211)H_(1915)N_(313)O_(364)S_(9),负电荷残基(Asp+Glu)总电荷为34,正电荷残基(Arg+Lys)总电荷为26,半衰期和脂肪指数分别为30 h和97.51。该蛋白的平均亲水性为-0.098,预测不稳定系数为27.13,故该蛋白属于亲水稳定蛋白,N端氨基酸为蛋氨酸(Met,M)。AcinJHBP在N端含一个长为17个氨基酸残基的信号肽,但无跨膜区;磷酸位点检测分别发现丝氨酸、苏氨酸和酪氨酸磷酸位点分别为9、6和3个,未发现糖基化位点。亚细胞定位结果显示,其主要位于细胞外。系统进化结果表明,AcinJHBP与同为鳞翅目尺蛾科的冬尺蠖(Operophtera brumata)血淋巴保幼激素结合蛋白JHBP亲缘关系最近。基因表达结果显示,在不同发育阶段,AcinJHBP基因在春尺蠖卵至5龄幼虫期间表达均较低(3龄幼虫除外),且无显著差异(P>0.05),而在非滞育蛹-成虫期,AcinJHBP表达量呈逐渐下降趋势。在不同组织中,春尺蠖AcinJHBP基因在雄虫头部表达量最高。[结论]AcinJHBP基因与春尺蠖生长发育与滞育密切相关,该结果为进一步揭示该基因在春尺蠖蛹越夏、越冬滞育中的功能奠定一定理论基础。[Objective]Thepresentpaperaimed to exploretheroleofAcinJHBP geneinthegrowth,developmentand diapauseofApocheima cinerarius.[Method]Based on the transcriptome data(non-diapause pupa/diapause pupa and diapause pupa/diapause depupa),JHBP gene was cloned and named as AcinJHBP(GenBank entry number:OR860357).The sequence and physicochemical properties of AcinJHBP gene were analyzed by bioinformatics software.The temporal and spatial expression of AcinJHBP(different developmental stages and tissues)was detected by qPCR.[Result]The complete CDS sequence of AcinJHBP gene of A.cinerarius was 726 bp,encoding 241 amino acids,the pre-dicted molecular weight of protein was 26.97 kDa,and the predicted isoelectric point was 5.13.The predicted molecular formula of AcinJH-BP gene was C_(1211)H_(1915)N_(313)O_(364)S_(9),the total charge of negative charge residues(Asp+Glu)was 34,the total charge of positive charge res-idues(Arg+Lys)was 26,the half-life and aliphatic index were 30 hours and 97.51,respectively.The average hydrophilicity of the protein was-0.098,and the predicted instability coefficient was 27.13.Therefore,the protein was a hydrophilic stable protein,and the N-termi-nal amino acid was methionine(Met,M).AcinJHBP contained a signal peptide of 17 amino acid residues at the N-terminus,but had no transmembrane region.Phosphoric acid sites were found in serine,threonine and tyrosine phosphate sites were 9,6 and 3,respectively,and no glycosylation sites were found.Subcellular localization results showed that it was mainly located outside the cell.Phylogenetic analysis showed that AcinJHBP had the closest relationship with the hemolymph juvenile hormone binding protein JHBP of Operophtera brumata.The results of gene expression showed that the expression of AcinJHBP was lower at different developmental stages(except the 3rd instar larvae),and there was no significant difference between the egg and the 5^(th)instarlarvae(P>0.05),for all that the expression of AcinJHBP showed a decreasing trend in non-diapause pupa-adult stage.In

关 键 词：春尺蠖 保幼激素结合蛋白基因 分子特征 表达谱 

分 类 号：S888.721[农业科学—特种经济动物饲养]