Analysis of SFL1 and SFL2 Promoter Region in Arabidipsis thaliana using Gateway Cloning System

SFL1 and SFl2 (SETH Four Like) genes are two members of SETH4 gene family in Arabidopsis thaliana expressed in saprophytic tissues. In this study, expression of SFL1 and SFL2 genes were studied using Gateway Cloning Technology. Primers were designed for PCR amplification of promoter region of SFL1 (900 bp) and SFL2 (930 bp) genes having attB1 recombination sites using Kod Hi Fi DNA polymerase enzyme. Amplified fragments were cloned into pDON201 vector by BP reaction and then into destination vector pGKWFS7 containing GFP::GUS by LR reaction. Finally, pGKWFS7-SFL1 and pGKWFS7-SFL2 were introduced into Agrobacterium GV3101 strain as a binary vector. Plants then were transformed with the new constructs. GUS staining of transgenic plants with SFL1 promoter showed strong expression in seedling, stem, leaf, root, root hair, sepal, sillique and unevenly staining in petal. Plants transformed with SFL2 showed the same pattern of expression (except root and root hair) but relatively weaker than SFL1. No expression of either SFL1 or SFL2 was observed in pollen.