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【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。
【Objective】 Metal-responsive transcription factor-1 (MTF-1) is a multifunctional transcription regulatory protein that plays a role in the body’s response to heavy metals, hypoxia or oxidative stress. This study aimed to determine the expression pattern of Oxya chinensis MTF-1 gene in different tissues and different developmental stages. 【Method】 The MTF-1 cDNA fragment was searched based on the Chinese locust transcriptome database. The 5 ’and 3’ end of MTF-1 was cloned by RACE method. Using different bioinformatics software, the resulting sequences were predicted by domain prediction, amino acid sequence alignment and phylogenetic tree analysis. According to the nucleotide sequence of the gene, specific expression primers were designed and their expression levels in different tissues and stages of Oxya chinensis were studied by RT-qPCR. 【Result】 The full-length cDNA sequence of MTF-1 from Oryxiasis japonicus was cloned and named as OcMTF-1 (GenBank accession number: KM017748), which was 1 834 bp in length with an open reading frame (ORF) of 1 527 bp encoding a 508 amino acids. Domain prediction showed that the sequence contained only 6 C2H2-type zinc finger structures, no other activation domain. Amino acid sequence alignment revealed that the zinc finger domain at the N-terminus of OcMTF-1 has a high similarity with MTF-1 of other insects known to date. OcMTF-1 was highly expressed in Markovichoderm of Chinese grasshopper, followed by hindgut and fat body, while the expression of OcMTF-1 in other tissues was relatively low. 【Conclusion】 The full-length cDNA of OcMTF-1 gene of O. chinensis was cloned in this study, and its zinc finger domain at the N-terminal of the protein has high identity with MTF-1 of other insects. The gene was highly expressed in M. asiatica, and was expressed in all developmental stages of nymphs and adults, with the highest expression in the first instar nymphs. The results provide the basic information for further exploring the transcriptional regulation mechanism of OcMTF-1 gene.