›› 2014, Vol. 57 ›› Issue (9): 1001-1007.

• RESEARCH PAPERS •     Next Articles

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

REN Li-Xian1, YU Zhi-Tao1, LIU Xiao-Jian1, ZHANG Jian-Zhen1, MA En-Bo1,*, GUO Ya-Ping2,*   

  1. (1. Institute of Applied Biology, Shanxi University, Taiyuan 030006, China;   2. College of Life Science, Shanxi University, Taiyuan 030006, China)
  • Online:2014-09-20 Published:2014-09-20

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