In order to set up a system for production of recombinant proteins using suspension cell culture of Nicotiana tabacum cv. Bright Yellow 2 (BY-2), in vitro culture and genetic transformation protocols of such cells were optimised at the laboratory of Plant Cell Biotechnology, Institute of Biotechnology. | JOURNAL OF SCIENCE OF HNUE Chemical and Biological Sci. 2012 Vol. 57 No. 8 pp. 128-137 This paper is available online at http OPTIMISING IN VITRO CULTURE AND Agrobacterium tumefaciens-MEDIATED TRANSFORMATION PROTOCOLS OF TOBACCO BY-2 CELLS Nguyen Tuong Van1 Le Quynh Lien1 and Nguyen Thanh Van2 1 Institute of Biotechnology Vietnam Academy of Science and Technology 2 Faculty of Biology Hanoi National University of Education Abstract. In order to set up a system for production of recombinant proteins using suspension cell culture of Nicotiana tabacum cv. Bright Yellow 2 BY-2 in vitro culture and genetic transformation protocols of such cells were optimised at the laboratory of Plant Cell Biotechnology Institute of Biotechnology. Our experiments indicated that the development rate of BY-2 cells depended on initial cell concentrations. The best culture condition was the formula F5 with 1 20 dilution at the starting point mL of initial cells 50 mL culture medium which reached the exponential phase after 5 days and had maximum biomass of g mL after 9 days. Agrobacterium tumefaciens-mediated transformation procedure of BY-2 cells was optimised by monitoring transient gusA gene expression. There is a positive correlation between the amount of BY-2 cells and the density of Agrobacterium tumefaciens in coculture medium. The transformation efficiency was the highest at formulars and bacteria OD-plant cell formular . Keywords BY-2 cells Agrobacterium tumefaciens-mediated transformation in vitro culture GUS transient expression coculture 1. Introduction Recombinant proteins are increasingly important components of medicine and applied chemistry. They are needed for a vast range of applications including therapeutics vaccines diagnostics and enzymes. Currently most recombinant proteins originate from genetically engineered bacteria. Other sources are eukaryotes like yeast human or animal cell lines or even transgenic animals. Compared
