中华稻蝗金属应答转录因子基因OcMTF-1的分子特性及表达分析

摘要/Abstract

摘要： 【目的】金属应答转录因子-1（metal responsive transcription factor-1, MTF-1）是一种多功能转录调控蛋白，可在机体应对重金属、低氧或氧化压力环境时发挥作用。本研究旨在确定中华稻蝗Oxya chinensis MTF-1基因在不同组织部位及不同发育时期的表达模式。【方法】基于中华稻蝗转录组数据库，搜索到MTF-1 cDNA片段，采用RACE方法克隆获得MTF-1的5′端和3′端序列。利用不同生物信息学软件对所得序列进行结构域预测、氨基酸序列比对和进化树分析。根据该基因的核苷酸序列，设计特异性表达引物，利用RT-qPCR方法研究其在中华稻蝗不同组织部位及不同发育时期的表达。【结果】克隆获得中华稻蝗MTF-1 cDNA全长序列，命名为OcMTF-1（GenBank登录号：KM017748），其长为1 834 bp，其中开放式阅读框（ORF）为1 527 bp，编码508个氨基酸。结构域预测发现，该序列只含有6个C2H2型锌指结构，无其他激活域。氨基酸序列比对发现，OcMTF-1 N端的锌指结构域与目前已知的其他昆虫的MTF-1有很高的相似度。OcMTF-1在中华稻蝗马氏管中高表达，其次是后肠和脂肪体，其他组织部位表达量相对较低；在1-5龄若虫期和成虫期均表达。【结论】本研究克隆获得中华稻蝗OcMTF-1基因cDNA全长序列，其编码蛋白N端的锌指结构域与其他昆虫MTF-1序列一致性较高。该基因在中华稻蝗马氏管中高表达；在若虫和成虫各发育阶段均有表达，其中1龄若虫期表达量最高。研究结果为进一步探索OcMTF-1基因的转录调控机制提供了基础资料。

关键词: 中华稻蝗, 金属应答转录因子-1, 基因克隆, RT-qPCR, 表达模式

Abstract: 【Aim】 The metal responsive transcription factor-1 (MTF-1) is a pluripotent transcriptional regulator, which plays an important role in the body response to heavy metals and hypoxia or oxidative stress. This study aims to determine the expression profiles of MTF-1 gene in different tissues and developmental stages of the Chinese rice grasshopper, Oxya chinensis. 【Methods】 The cDNA fragment of MTF-1 was obtained from the transcriptome database of O. chinensis, and its 5′ and 3′ sequences were cloned using RACE methods. The structure prediction, sequence alignment and phylogenetic analysis of the coding product of this gene were performed using different bioinformatics softwares. Based on the cloned mRNA nucleotide sequences of MTF-1, specific primers were designed to analyze its developmental and tissuespecific expression patterns using RT-qPCR. 【Results】 The full length cDNA of MTF-1 from O. chinensis was cloned and named OcMTF-1 (GenBank number: KM017748). Its length is 1 834 bp, with a 1 527 bp open reading frame encoding a 508amino acid polypeptide. It has six C2H2- type zinc finger regions only and no other activating domains were identified. Multiple alignments revealed that the zinc finger domain is conserved at the N terminus of OcMTF-1 and MTF-1 proteins from other species. RT-qPCR analysis showed that the expression level of OcMTF-1 was the highest in the Malpighian tubules, moderate in the hindgut and fat body, and the lowest in other tissues. OcMTF-1 was expressed at both the 1st to 5th instar nymphal stages and adult stage. 【Conclusion】 In this study, we cloned a full length sequence of OcMTF-1 cDNA, and the zinc finger domain at the N terminus of OcMTF-1 has high sequence identity with MTF-1 proteins from other insects. OcMTF-1 was highest expressed in the Malpighian tubules, and expressed in all the developmental stages with the highest expression level in the 1st instar nymph. The results provided a foundation for further research on the mechanism of gene regulation of OcMTF-1.

Key words: Oxya chinensis, metal responsive transcription factor-1 (MTF-1), gene cloning, RT-qPCR, expression pattern

任丽娴，余志涛，刘晓健，张建珍，马恩波，郭亚平. 中华稻蝗金属应答转录因子基因OcMTF-1的分子特性及表达分析[J]. , 2014, 57(9): 1001-1007.

REN Li-Xian, YU Zhi-Tao, LIU Xiao-Jian, ZHANG Jian-Zhen, MA En-Bo, GUO Ya-Ping. Molecular characterization and expression analysis of metal responsive transcription factor-1 gene OcMTF-1 from Oxya chinensis (Orthoptera: Acridoidea)[J]. , 2014, 57(9): 1001-1007.

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中华稻蝗金属应答转录因子基因OcMTF-1的分子特性及表达分析

Molecular characterization and expression analysis of metal responsive transcription factor-1 gene OcMTF-1 from Oxya chinensis (Orthoptera: Acridoidea)

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