development of useful recombinant promoter and its expression analysis in different plant cells using confocal laser scanning microscopy开发有用的启动子及其重组表达分析在不同植物细胞使用共焦激光扫描显微镜.pdfVIP

Development of Useful Recombinant Promoter and Its Expression Analysis in Different Plant Cells Using Confocal Laser Scanning Microscopy 1 1. 1. 1,2 2 1 Deepak Kumar , Sunita Patro , Rajiv Ranjan , Dipak K. Sahoo , Indu B. Maiti , Nrisingha Dey * 1 Department of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekherpur, Bhubaneswar, Orissa, India, 2 Kentucky Tobacco Research and Development Center (KTRDC), College of Agriculture, University of Kentucky, Lexington, Kentucky, United States of America Abstract Background: Designing functionally efficient recombinant promoters having reduced sequence homology and enhanced promoter activity will be an important step toward successful stacking or pyramiding of genes in a plant cell for developing transgenic plants expressing desired traits(s). Also basic knowledge regarding plant cell specific expression of a transgene under control of a promoter is crucial to assess the promoter’s efficacy. Methodology/Principal Findings: We have constructed a set of 10 recombinant promoters incorporating different up- stream activation sequences (UAS) of Mirabilis mosaic virus sub-genomic transcript (MS8, -306 to +27) and TATA containing core domains of Figwort mosaic virus sub-genomic transcript promoter (FS3, 2271 to +31). Efficacies of recombinant promoters coupled to GUS and GFP reporter genes were tested in tobacco protoplasts. Among these, a 369-bp long hybrid sub-genomic transcript promoter (MSgt-FSgt) showed the highest activity in both transient and transgenic systems. In a transient system, MSgt-FSgt was 10.31, 2.86 and 2.18 times