沙葱萤叶甲GdHsp70-2,GdHsp70-3的克隆鉴定与表达谱分析  

Cloning,Identification and Expression Profiling Analysis of GdHsp70-2,GdHsp70-3 Genes in Galeruca daurica

作　　者：张宏玲 任浩 田羽 单艳敏 王莹 李玲 李艳艳 庞保平 谭瑶 ZHANG Hong-ling;REN Hao;TIAN Yu;SHAN Yan-min;WANG Ying;LI Ling;LI Yan-yan;PANG Bao-ping;TAN Yao(Research Center for Grassland Entomology,Inner Mongolia Agricultural University,Hohhot,Inner Mongolia 010019,China;Grassland Station of Bordered Yellow Banner,Bordered Yellow Banner,Inner Mongolia 013250,China;Inner Mongolia Forestry and Grassland Pest Control and Qurantine,Hohhot,Inner Mongolia 010051,China;Ordos Natural Resources Bureau Kangbashi Branch,Ordos,Inner Mongolia 017010,China)

机构地区：[1]内蒙古农业大学草原昆虫研究中心,内蒙古呼和浩特010019 [2]镶黄旗草原工作站,内蒙古镶黄旗013250 [3]内蒙古自治区林业和草原有害生物防治检疫总站,内蒙古呼和浩特010051 [4]鄂尔多斯市自然资源局康巴什区分局,内蒙古鄂尔多斯017010

出　　处：《草地学报》2023年第1期61-72,共12页Acta Agrestia Sinica

基　　金：内蒙古高等学校青年科技英才计划(NJYT-19-B22);中国博士后科研启动项目(155582);内蒙古自治区关键技术攻关项目(2021GG0108)资助。

摘　　要：为明确沙葱萤叶甲Galeruca daurica热激蛋白70(Heat shock protein 70,Hsp70)基因的序列结构及其系统进化关系,本研究通过PCR技术克隆沙葱萤叶甲Hsp70基因cDNA全长序列与基因组序列,并进行生物信息学与表达谱分析。结果显示,克隆获得了沙葱萤叶甲2条Hsp70基因GdHsp70-2(GenBank登录号:MZ853083)和GdHsp70-3(Genbank登录号:OK585088),基因全长分别为2410 bp和2242 bp,各自编码657和646个氨基酸,均含有3个保守的HSP70家族特征序列,预测蛋白三维结构均由N-端ATPase功能域和C-端底物结合功能域所组成;系统发育分析表明GdHSP70-2,GdHSP70-3分别与松墨天牛(Monochamus alternatus)MaltHSC70-1、玉米根萤叶甲(Diabrotica virgifera virgifera)DvirHSP70-2的亲缘关系最近;基因组DNA克隆获得GdHsp70-2的两段内含子序列,GdHsp70-3则不含有内含子;表达谱分析结果表明GdHsp70-2,GdHsp70-3均在蛹期的表达量最高,在成虫中分别在翅和足的表达量最高。研究有助于加深对沙葱萤叶甲Hsp70基因结构及其进化关系的了解,并为后续深入开展功能研究提供理论依据。To identify the gene structure of the heat shock protein 70(Hsp70)of Galeruca daurica and further explore their phylogenetic relationship.Polymerase chain reaction(PCR)was used to clone the full-length cDNA and genomic sequence of Hsp70 gene from G.daurica.The sequence was subjected to bioinformatical and expression patterns analysis.The results showed that the two complete cDNA sequences were obtained and named GdHsp70-2(GenBank accession No.:MZ853083)and GdHsp70-3(GenBank accession No.:OK585088).Their full-length cDNA were 2410 and 2242 bp in length,encoding 657 and 646 amino acids,containing three highly conserved heat shock protein 70 family regions,and the 3D structure of the two HSP70 were composed of the N-terminal ATPase functional domain and C-terminal substrate binding domain,respectively.The phylogenetic tree showed that GdHSP70-2 and GdHSP70-3 have the highest amino acid sequence identity with MaltHSC70-1 of Monochamus alternatus,and DvirHSP70-2 of Diabrotica virgifera virgifera.The genomic sequence cloning results displayed that GdHsp70-2 has two introns,and the GdHsp70-3 contains no intron in the gene.Expression profile analysis showed that the expression levels of GdHsp70-2 and GdHsp70-3 in pupae were significantly higher than that in other developmental stages,and the relative expression level in wings and feet was the highest in adults.These findings improve the understanding of gene structure and its evolutionary relationships of G.daurica HSP70 family genes and provide a theoretical basis for gene function studies.

关 键 词：沙葱萤叶甲 热激蛋白70 基因克隆 表达谱分析 内含子 

分 类 号：Q966[生物学—昆虫学]