Cloning,expression analysis and autonomous transcriptional activation testing of transcription factor LeAGL11 from Lagerstroemia excelsa(Dode)Chun
[Objective]To investigate the differential expression of AGL11 in self-and cross pollinations between Lagerstroemia indica and Lagerstroemia excelsa(Dode)Chun,and self-activation characteristics of the transcription factor AGL11,it can lay a foundation for the study of the biological function of LeAGL11 in the distant cross incompatibility between L.excelsa and L.indica.[Method]We cloned the LeAGL11 and analyzed its physicochemical property through bioinformatics,RT-qPCR was used to assess and compare the expression of LeAGL11 in pistils of different developmental stages(24 h,48 h and 72 h after'Ziyun'×'Ziyun'and'Ziyun'×'L.excelsa No.1'),a pGBKT7-AGL11 yeast expression vector was constructed by DNA recombination technology and transformed into the Y2HGold yeast strain.[Result]The coding sequence of LeAGL11 was 666 bp,translating 221 amino acids,it had no signal peptide and transmembrane domain,it was hydrophilic protein but not membrane protein,the sequence alignments of amino acids and phylogenetic analysis showed that it had a high degree of similarity with the AGL11 protein of L.indica,Punica granatum and Vitis vinifera.The protein structure prediction and sequence analysis indicated that it had a typical MADS-box and K-box conserved domains of the MADS-box gene family.The relative expression of this gene was highest expressed at 24 h after self-pollination,showing an upward trend in selfing,and firstly decreased and then increased up in hybridization.Self-activation showed pGBKT7-LeAGL11 plasmid did not turn blue in SD/-Trp+AbA+X-α-gal medium.[Conclusion]The relative expression of the LeAGL11 gene during selfing in L.indica and hybridization between L.indica and L.excelsa is characterized by a certain degree of variability,the LeAGL11 gene has no self-activation activity and could be used for subsequent yeast two hybrid experiments.
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