Cloning the RGL2 gene from Arabidopsis thaliana genomic DNA

Document Type : Research Paper

Author

Abstract

There are vast endeavors by scientists to investigate gibberellin synthesis cycle in plant kingdom. One of the known genes that are supposed to be effective in gibberellin synthesis is RGL2 which was cloned in this study.  Using extracted DNA from Colombia ecotype of Arabidopsis thaliana, the gene was amplified using specific primers designed based on the RGL2 gene sequence. The PET-32a(+) plasmid was selected for the gene cloning. The plasmid was digested by two specific restriction enzymes, Nco I and Sal I along with the DNA sample. Ligating the gene to the vector, an E. coli strain, XAL-Gold was used for the gene transformation. PCR conducted on the transformed bacteria confirmed the accuracy of the cloning.

Keywords

Main Subjects

References

Berardini, T.Z., Bollman, K., Sun, H., and Poethig, R.S., 2001. Regulation of vegetative phase change in Arabidopsis thaliana by cyclophilin 40. Science, 291: 2405–2407.
Bewley, J.D., 1997. Seed germination and dormancy. Plant Cell, 9: 1055-1066.
Davies, P.J., 1995. Plant Hormones: Physiology, Biochemistry, and Molecular Biology. Kluwer Academic Publishers, Dordrecht, the Netherlands.
Dill, A., Jung, H.S., and Sun, T.P., 2001. The DELLA motif is essential for gibberellin-induced degradation of RGA. Proc. Natl. Acad. Sci. USA, 98: 14162–14167.
Fleet, C.M. and Sun, T.P., 2005. A DELLAcate balance: the role of gibberellin in plant morphogenesis. Curr. Opin. Plant Biol., 8: 77–85.
Gan, Y., Kumimoto, R., Liu, C., Ratcliffe, O., Yu H. and Broun, P., 2006. Glabrous inflorescence stems modulates the regulation by gibberellins of epidermal differentiation and shoot maturation in Arabidopsis. The Plant Cell, 18: 1383-1395.
Groot, E.P. and Meicenheimer, R.D., 2000. Comparison of leaf plastochron index and allometric analyses of tooth development in Arabidopsis thaliana. J. Plant Growth Regul., 19: 77–89.
Hunter, C., Sun, H. and Poethig, R.S., 2003. The Arabidopsis heterochronic gene ZIPPY is an ARGONAUTE family member. Curr. Biol., 13: 1734–1739.
Lee, S., Cheng, H., King, K. E., Wang, W., He, Y., Hussain, A., Lo, J., Harberd, N. P. and Peng, J., 2002. Gibberellin regulates Arabidopsis seed germination via RGL2, a GAI/RGA-like gene whose expression is up-regulated following imbibitions Genes and Development, 16: 646-658,
McGinnis, K.M., Thomas, S.G., Soule, J.D., Strader, L.C., Zale, J.M., Sun, T.P. and Steber, C.M., 2003. The Arabidopsis SLEEPY1 gene encodes a putative F-box subunit of an SCF E3 ubiquitin ligase. Plant Cell, 15: 1120–1130
Meyerowitz, E.M. and Somerville, C.R., 1994. Arabidopsis. Cold Spring Harbor Laboratory Press, NY.
Murray, M.G. and Thompson, M.F., 1980. Rapid isolation of high molecular weight plant DNA. Nucl. Acid Res. 8: 4321-4325.
Nakajima, K., Sena, G., Nawy, T. and Benfey, P.N., 2001. Intercellular movement of the putative transcription factor SHR in root patterning. Nature, 413: 307-311.
Peng, J.R., Carol, P., Richards, D.E., King, K.E., Cowling, R.J., Murphy, G.P. and Harberd, N.P., 1997. The Arabidopsis GAI gene defines a signaling pathway that negatively regulates gibberellin responses. Genes and Development. 11: 3194-3205.
Peng, J.R., Richards, D.E., Hartley, N.M., Murphy, G.P., Devos, K.M., Flintham, J.E., Beales, J., Fish, L.J., Worland, A.J. and Pelica, F., 1999. "Green Revolution" genes encode mutant gibberellin response modulators. Nature 400: 256-261.
Silverstone, A.L., Ciampaglio, C.N. and Sun, T.P., 1998. The Arabidopsis RGA gene encodes a transcriptional regulator repressing the gibberellin signal transduction pathway. Plant Cell 10: 155-169.
Steber, C.M, Cooney, S.E., and McCourt, P., 1998. Isolation of the GA-response mutant sly1 as a suppressor of ABI1-1 in Arabidopsis thaliana. Genetics 149: 509-521.
Iranian Journal of Rangelands and Forests Plant Breeding and Genetic Research
Volume 15, Issue 4 - Serial Number 4
February 2008
Pages 265-272

Files

Share

How to cite

Statistics