| Promoters are one of the important factors that regulated gene expression.Althouhg the understanding of these processes has significant agronomie value，relatively few studies of regulation of these poreesess have been peorfmred. PlantWUS gene-encoded transcriptional regulators appear to be made the surrounding cellswith the characteristics of stem cell. In the way of maintaining its central region to thenumber of stem cell populations and specificity to improve the meristem activity ofstems and flowers. Therefore, screening the WUS gene promoter is very significant ofMaize.We clone three WUS gene promoters in Zea Maize, and analyse the functionswith GUS reporter gene. Through the promoter reduction experiment, rice genetictransformation and GUS staining, we get the acheievements as below:1. Using bioinformatics software, a total of19genes had been found to encodeputative WUS genes and their chromosomal locations, phylogenetic tree. Analysingthe upstream promoter fragments of the highly homologous genes based on thehomology results.2. The promoters fragment of WUS gene are amplified by polymerase chainreaction (PCR) technique from the DNA of maize B73, we called them ZmPw1、ZmPw3、ZmPw5. We forecast some cis-elements in the promoters from PLACEwebsite.3. Besides upstream promoter of WUS1gene, five5’missing promoters arecloned, whose length are1375bp、1095bp、807bp、538bp and171bp respectively. Thethree promoters and plasmid pCAMBIA1301which contains GUS reporter gene areattached to construct pZmPw1、pZmPw3、pZmPw5and five5’ missing promotercarriers whose name are pZmPw1-2、pZmPw1-3、pZmPw1-4、pZmPw1-5、pZmPw1-6respectively.4. Transferring all plant expressional vectors to rice byAgrobacterium-mediated transformation,13transgenic plants are obtained. Accordingto experimental results, the GUS staining result of root, stipule, stem, leave related topZmPw1-5vector is blue. In conclusion, the core area of ZmPw1promoter is between538bp and0bp before transcription start site. |
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