Development of Transient Gene Expression System using Seedlings

다국어 초록 (Multilingual Abstract)

An efficient transient expression system has been developed and characterized for the production of foreign genes in seedlings. The seedlings can be easily produced from commercial seeds used for vegetable sprouts. In principal, a chemical abrasive was employed to generate wounds in seedlings prior to vacuum-infiltration with Agrobacterium tumefaciens bearing the target gene. This optimized chemical wounding-assisted agro-infiltration process resulted in up to 15-fold increase in β-glucuronidase (GUS) enzyme activity. This procedure has been used efficiently to express hepatitis B surface antigen (HBsAg) protein in a transient mode. Therefore, seedlings with proper wounds can be suggested as a convenient tool for the production of useful recombinant proteins.

참고문헌 (Reference)

1 Singh, N., "Use of silicon carbide fibers for Agrobacterium-mediated transformation in wheat" 76 : 1483-1485, 1999

2 Kim, S. S., "Transient β-glucuronidase expression in lily (Lilium longiflorum L.) pollen via wounding-assisted Agrobacteriummediated transformation" 29 : 965-969, 2007

3 Tiollais, P., "The hepatitis B virus" 317 : 489-495, 1985

4 Samac, D. A., "Strain specificity in transformation of alfalfa by Agrobacterium tumefaciens" 43 : 271-277, 1995

5 Murray, M. G., "Rapid isolation of high molecular weight plant DNA" 8 : 4321-4325, 1980

6 Tiwari, S., "Plants as bioreactors for the production of vaccine antigens" 27 : 449-467, 2009

7 Smith, M. L., "Hepatitis B surface antigen (HBsAg) expression in plant cell culture: Kinetics of antigen accumulation in batch culture and its intracellular form" 80 : 812-822, 2002

8 Lee, Y. W., "Genetic evidence for direct sensing of phenolic compounds by the VirA protein of Agrobacterium tumefaciens" 92 : 12245-12249, 1995

9 Mason, H. S., "Expression of hepatitis B surface antigen in transgenic plants" 89 : 11745-11749, 1992

10 Hadiji-Abbes,N.,I.Borchani-Chabchoub,H.Triki,R.Ellouz,A.Gargouri,and R.Mokdad-Gargouri, "Expression of HBsAg and preS2-S protein in different yeast based system: A comparative analysis" 66 : 131-137, 2009

11 Zhang, H., "Development of alfalfa (Medicago sativa L.) regeneration system and Agrobacterium-mediated genetic transformation" 9 : 170-178, 2010

12 Floros, J. D., "Chemical (NaOH) peeling as viewed by scanning electron microscopy: pimiento peppers as a case study" 52 : 1312-1316, 1987

13 Rakoczy-Trojanowska, M., "Cell. Mol. Biol. Lett." 7 : 849-858, 2002

14 Renard, C. M. G. C., "Bleaching of apple pomace by hydrogen peroxide in alkaline conditions: Optimisation and characterization of the products" 30 : 398-405, 1996

15 Jefferson, R. A., "Assaying chimeric genes in plants: the gus gene fusion system" 5 : 387-405, 1987

16 Streatfield, S. J., "Approaches to achieve high-level heterologous protein production in plants" 5 : 2-15, 2007

17 Flores Solís, J. I., "Application of sonication-assisted Agrobacterium-mediated transformation in Chenopodium rubrum L" 49 : 255-260, 2003

18 Opabode, J. T., "Agrobacterium-mediated transformation of plants: emerging factors that influence efficiency" 1 : 12-20, 2006