Cloning And Functional Analysis Of WRKY Gene Promoter In Zea Mays

Promoters are important factors that regulated gene expression. The activity of promoters indirectly reflect gene expression that they control. As one kind of promoter, tissue-specific promoters could drive stable and effective expression of target gene in specific tissues of plants. As a result, wastes are reduced. WRKY gene family is a type of transcriptional regulatory factors which has specific functions in plants. Plant WRKY gene-encoded trans criptional regulators appear to be involved in various physiological programs, including disease-resistance, senescence, stress responses of biotic and abiotic, growth and development processes.We clones a WRKY gene promoter in Zea Maize, and analyse the functions with GUS reporter gene. Through the promoter reduction experiment, rice genetic transformation and GUS staining, we get the acheievements as below:1. A2880bp promoter fragment of WRKY gene is amplified by polymerase chain reaction (PCR) technique from maize B73, we called it P2880. We forecast some cis-elements in P2880from PLACE website.2. Besides upstream promoter of WRKY gene, four5’missing promoters are cloned, whose length are2880bp,1812bp,1254bp,680bp and355bp respectively. These five promoters and plasmid pCAMBIA1301which contains GUS reporter gene are attached to construct pCAM2880GUS and four5’missing promoter carriers whose name are pCAM1812GUS, pCAM1254GUS, pCAM680GUS and pCAM355GUS respectively.3. Transferring all plant expressional vectors to rice by Agrobacterium-mediated transformation,37transgenic plants are obtained. According to experimental results, the GUS staining result of callus tissue related to pCAM2880GUS vector is blue, whereas, other vectors such as pCAM1812GUS, pCAM1254GUS, pCAM680GUS and pCAM355GUS are diffrent. In conclusion, the core area of P2880promoter is between2880bp and1812bp before transcription start site.To sum up, we clone a WRKY gene promoter in maize, and found that the promoter is a callus-specific promoter. There is no report of WRKY gene which is callus-specific. The cloning of this promoter has the vital significance.