Poster: Education

Abs # 25: Direct DNA transformation into plant by positively charged oligopeptides

Presenter:	Bong Gu, Kang , inpains@empal.com
Authors	Bong Gu, Kang  (A)   Woo Sung, Lee  (A)
Affiliations:	(A): Bong Gu Kang ,Young Sun Oh, Jung A Son, Woo Sung Lee, Department of Biological Sciences,Sungkyunkwan University

Abstract Oligopeptide-mediated gene transformation was shown to be successful in transforming mammalian cells. This method involves electrostatic interaction between oligopeptide and DNA, penetration of cell membrane, and nucleus targeting. In this study, polylysine and polyarginine oligopeptides were successfully employed to deliver and express green fluorescent protein (GFP) or b-glucuronidase (GUS) gene under the control of 35S promoter in tobacco suspension cell or selected tissues of onion and water celery[Onenanthe javanica(Blume)DC]. Prior to 'in planta transformation' we carried out gene delivery experiments in tobacco suspension cell. GFP or GUS expression was also observed in tobacco suspension cells when cells were incubated in the presence of the complex. These peptide/DNA complexes in buffer solution were incubated with intact tissues of the onion for 1-3 h and these tissues grew further for various intervals in the absence of the complex. GFP expression was observed in approximately 50% of onion or water celery root cells examined at the third day of the further growth. After the two weeks of further growth, still approximately 50 % of cells in the further-grown regions showed GFP expression, indicating that GFP gene was integrated into chromosome and was passed to the daughter cells. It was also shown that these oligopeptides were able to effectively target nucleus. This property of these oligopeptides was thought to be a major cause for the efficient GFP expression.