Transformasi Genetik Tembakau dengan Gen Cold Shock Protein melalui Perantara Agrobacterium tumefaciens

Abstract

Cold shock protein (Csp) essential for organisms to survive in abiotic stress condition. CspB gene has been fused to ubiquitin promoter in the T-DNA region of pCambia 1300int, and introduced into Agrobacterium tumefaciens LBA4404. This research had an objective to transform genetically Nicotiana tabacum cv. Samsun by CspB gene under the control of Ubiquitin promoter and NOS terminator mediated by A. tumefaciens. Leaf discs were co-cultivated with A. tumefaciens LBA 4404. Based on the number of hygromycinresistant calli, the efficiency of transformation was 57.5%. In the selective medium containing 50 μg/l hygromycin, the efficiency of regeneration of transgenic shoots was 82.6%. Based on PCR analysis using primers corresponding to ubiquitin promoter and CspB gene, 18 putative tobacco transgenic containing CspB gene under the control of ubiquitin promoter.