Development of a salicylic acid inducible minimal sub-genomic transcript promoter from Figwort mosaic virus with enhanced root- and leaf-activity using TGACG motif rearrangement

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• 作者：Deepak Kumar^a ; ^b ; ^{dpkbiotech@gmail.com} ; Sunita Patro^a ; ^b ; ^{sunitapatro07@gmail.com} ; Jayasish Ghosh^a ; ^b ; ^{joy.biochem@gmail.com} ; Abhimanyu Das^a ; ^b ; ^{touchabhimanyu@gmail.com} ; Indu B. Maiti^c ; ^{imaiti@uky.edu} ; Nrisingha Dey^a ; ^b ; ^{dey@ils.res.in} ; ^{nrisinghad@yahoo.com}
• 关键词：FMV ; Figwort mosaic virus ; CaMV ; cauliflower mosaic virus ; ASF-1 ; activation sequence factor-1 ; as-1 ; activation sequence鈥? ; GUS ; 尾-glucuronidase ; EMSA ; electrophoretic mobility shift assay ; CLSM ; confocal laser scanning microscopy
• 刊名：Gene
• 出版年：2012
• 期刊代码：73_03781119
• 类别：bio
• 出版时间：15 July, 2012
• 卷：503
• 期：1
• 页码：36-47
• 文件大小：1273 K

摘要

In Figwort mosaic virus sub-genomic transcript promoter (F-Sgt), function of the TGACG-regulatory motif, was investigated in the background of artificially designed promoter sequences. The 131 bp (FS, 鈭?#xA0;100 to + 31) long F-Sgt promoter sequence containing one TGACG motif [FS-(TGACG)] was engineered to generate a set of three modified promoter constructs: [FS-(TGACG)₂, containing one additional TGACG motif at 7 nucleotides upstream of the original one], [FS-(TGACG)₃, containing two additional TGACG motifs at 7 nucleotides upstream and two nucleotides downstream of the original one] and [FS-(TGCTG)_mu, having a mutated TGACG motif]. EMSA and foot-printing analysis confirmed binding of tobacco nuclear factors with modified TGACG motif/s. The transcription-activation of the GUS gene by the TGACG motif/s in above promoter constructs was examined in transgenic tobacco and Arabidopsis plants and observed that the transcription activation was affected by the spacing/s and number/s of the TGACG motif/s. The FS-(TGACG)₂ promoter showed strongest root-activity compared to other modified and CaMV35S promoters. Also under salicylic acid (SA) stress, the leaf-activity of the said promoter was further enhanced. All above findings were confirmed by real-time and semi-qRT PCR analysis. Taken together, these results clearly demonstrated that the TGACG motif plays an important role in inducing the root-specific expression of the F-Sgt promoter. This study advocates the importance of genetic manipulation of functional cis-motif for amending the tissue specificity of a plant promoter. SA inducible FS-(TGACG)₂ promoter with enhanced activity could be a useful candidate promoter for developing plants with enhanced crop productivity.