Abstract
The first successful attempt to produce transgenic tobacco cell suspensions expressing a modified version of amarantin, through direct gene transfer using particle gun is described. The physical parameters for transformation were optimized using the gene encoding β-glucuronidase (gusA) as the reporter gene. Statistical analysis revealed that helium pressure, target distance, and osmotic pretreatment had significant influence on transformation efficiency. Of the different variables evaluated, a combination of sorbitol 0.5 M pretreatment, shooting distance of 7 cm and firing pressure of 900 psi were found ideal. Selection of putative transgenic calli was done on Hygromicine-selection medium. Presence of the transgene was confirmed using histological glucuronidase assay and PCR analysis. Accumulation and stable expression of the recombinant amarantin was detected using Western blot analysis. Transformation efficiency in terms of biochemical GUS assay was of 60%. The present results demonstrate the possibility of incorporating and expressing transgene encoding recombinant proteins of interest, in tobacco cell suspension through biolistic delivery system.
Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
