Bioinformatic and molecular analysis of seed-specific promoter SBP in Tobacco

Tissue-specific promoters limit gene expression to certain cells, tissues, organs, or developmental stages that have many applications in genetically modified crops[1]. Considering the importance of increasing seed oil content in safflower, in this study, a synthetic seed specific promoter, sucrose binding protein (SBP) from Vicia faba was tested for its activity in transgenic tobacco plants. The probably regulatory cis-elements of the promoter sequence were identified using the online software tools PLANTCARE[2] and PLACE[3]. Database searching revealed that this region contains eleven cis-acting regulatory element involved in seed-specific regulation which are CAATBOX1, ABREOSRAB21, DPBFCOREDCDC3, SEF1MOTIF, SEF4MOTIFGM7S, MYB1AT, -300ELEMENT, CAREOSREP1, GADOWNAT, GADOWNAT and CEREGLUBOX2PSLEGA. The SBP promoter sequence was synthesized by GENEray Company. Correctness of the promoter region was confirmed by restriction enzyme digestion and PCR. The sequence was inserted to the GUS gene upstream in pBI121 vector using general cloning methods[4] to produce SBP promoter or GUS construct. The construct was transformed into Agrobacterium tumefaciens strain EHA105[5] and then Tobacco plants were transformed by leaf disk procedures. The functionality of the SBP promoter was confirmed in stably transformed tobacco plants and it can be employed for seed specific expression in crop breeding projects

Keywords:

sucrose binding protein , tissue , specific promoter , tobacco , PLACE , PLANTCARE