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Development of Rescue Cloning Vector with Phosphinothricin Resistant Gene for Effective T-DNA Tagging  

Kim, Hyoung-Seok (Graduate School of Biotechnology & Department of Horticulture, KyungHee University)
Kim, Sung-Hoon (Graduate School of East-West Medical Science, KyungHee University)
Park, Young-Doo (Graduate School of Biotechnology & Department of Horticulture, KyungHee University)
Publication Information
Horticulture, Environment, and Biotechnology : HEB / v.44, no.4, 2003 , pp. 407-411 More about this Journal
Abstract
This study was carried out to develop an effective rescue cloning vector, pRCV3, for T-DNA tagging. In phosphinothricin (PPT) herbicide added medium, the newly developed vector pRCV3 possesed PPT resistant genes which were capable of selecting a characteristic transformation system. It also contained within T-DNA, ampicillin resistant gene and bacterial replication origin for accomplishing plasmid rescue. To isolate both left and right insert junctions of a T-DNA tag, one unique restriction enzyme site within the vector was removed. Moreover, to detect various unpredictable integration patterns, PCR confirmation primer sets, which can be amplified in several important regions of the vector, were prepared. Based on these data, sequencing primers were designed to use directly for analysis of flanking plant DNA. To confirm an applicability of developed rescue cloning vector, tobacco (Nicotiana tabaccum cv. Havana SR1) was transformed with Agrobacterium tumerfaciens LBA4404 harboring pRCV3, and then plasmid rescue technique was applied to transformants which were confirmed by PCR
Keywords
Agrobacterium tumefaciens; flanking plant DNA; integration pattern;
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1 Feldmann, K.A. 1992. p. 274-289. In: C. Koncz, H.H. Chua, and J. Schell (eds.). Methods in Arabidopsis research. World Scientific
2 Gustavo, A., J. Gonzalez-Cabrera, R. Vazquez-Padron, and C. Ayra-Pardo. 1998. Agrobacterium tumefaciens: A natural tool for plant transformation. Electronic J. Biotechnol. 1:1-16
3 Pansegrau, W. and E. Lanka. 1991. Common sequence motifs in DNA relaxases and nick region from a variety of DNA transfer system. Nucl. Acids Res. 19:3455   DOI   ScienceOn
4 Park, Y.D. and H.S. Kim. 2000. Expression and inheritance patterns of gus gene driven by an endosperm-specific promoter in transgenic tobacco. Kor. J. Hort. Sci. Technol. 18:594-598
5 Yanofsky, M.F., H. Ma, J.L. Bowman, G.N. Drews, K.A. Feldmann, and E.M. Meyerowitz. 1990. The protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors. Nature 346:35-39   DOI   ScienceOn
6 Elizabeth, E.H. 1999. Analysis of plant transformation system. The thirtieth anniversary seminar of the Korean society of breeding. p. 33-37
7 An, G., P.R. Ebert, A. Mitra, and S.B. Ha. 1988. Binary vectors, p. 1-19. In: S.B. Gelvin, R.A. Schilperoort, and D.P.S. Verma (eds.). Plant molecular biology mannual, A3. Academic Publishers, Dondrecht, Belgium
8 Martineau, B., T.A. Voelker, and R.A. Sanders. 1994. On defining T-DNA. 6:1032-1033
9 van der Graaff. E., A. den Dulk-Ras, and P.J.J. Hooykaas. 1996. Deviating T-DNA transfer from Agrobacterium tumefaciens to plants. Plant Mol. Biol. 31:667-681   DOI
10 Mathur, J., L. Szabados, S. Schaefer, B. Grunenberg, A. Lossow, E. Jonas-Straube, J. Schell, C. Koncz, and Z. Koncz-Kalman. 1998. Gene identification with sequenced T-DNA tags generated by transformation of Arabidopsis cell suspension. Plant J. 13:707-716   DOI   ScienceOn