Cloning and Sequence Analysis of SmWRKY1 Transcription Factor Gene in Eggplant

doi:10.11924/j.issn.1000-6850.2013-2505

Abstract

Abstract: The aims were to clone the transcription factor SmWRKY1 gene in eggplant and analyze its sequence characteristics, and provide a theoretical basis for the application of molecular breeding of SmWRKY1. Eggplant seedlings were used as experimental materials. Specific primers were designed based on the sequences of WRKY gene from tomato, sweet pepper and tobacco, which has been submitted to GenBank. SmWRKY1 gene fragment was amplified by RT- PCR. NCBI online and DNAMAN software were used to analyze nucleic acids and amino acid sequences. SmWRKY1 were cloned through homology-based cloning. SmWRKY1 included 930 nucleotide and encoding 310 amino acids, where Ser and Thr are largest amount of the total number of amino acids with 12.0% and 11.0%. NCBI BLAST showed that SmWRKY1 belonged to WRKY gene superfamily with 114th to 120th amino acids containing WRKY conserved domain‘WRKYGQK’and the zinc finger structure of type C2H2. SmWRKY1 was confirmed belonging to the second category of WRKY superfamily. Plant phylogenetic tree including SmWRKY1 proteins and other plant proteins was built through DNAMAN software, showed that SmWRKY1 had the closest relationship with tomato WRKY1 protein with homology of 96%. WRKY gene played an important role in plant stress tolerance and this research cloning gene SmWRKY1 fragment made early preparations for further cloning of its full-length and the studing of its functions and laid a certain foundation for molecular breeding and cultivation of new varieties of eggplant.

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