Keywords
E. coli
Bivalent salt
How to Cite
Abstract
Bacterial transformation is the process of up taking foreign DNA molecules by bacterial cells. Bacteria, which are able to up take such foreign DNA are called “competent” cell. Foreign DNA carrying antibiotic resistance gene can be expressed in the bacteria and transforming the bacteria from antibiotic sensitive to antibiotic resistance. Such transformed bacteria can easily be isolated. The efficiency of the transformation plays a critical role in recombinant DNA technology. Several methods have been developed to improve the transformation efficiency. In our work we have studied the transformation efficiency on strains of E. coli and in response to different bivalents salts at different concentrations. All the experiments revealed that transformation efficiency was highest on E. coli DH5α strain in 50 mM CaCl2 concentration. Comparing to the other E. coli and bivalents salts concentration, 50 mM CaCl2, resulted in much faster growth of transformed bacteria. Thus, our results suggest that E. coli DH5α strain and CaCl2 can be used as an effective agent for the transformation technique.
References
Brian P and Heller Mk. 1996. High efficiency 5 min transformation of Escherichia coli. Nucleic Acids Research 24(3):536-537.
Hanahan D, Jessee J, Bloom FR. 1991. Plasmid transformation of Escherichia coli and other bacteria. Methods in Enzymology. 204:63-113.
Hanahan D. 1983. Studies on transformation of E.coli with plasmids. Journal of Molecular Biology. 166:557-580.
Mandel M and Higa A. 1970. Calcium dependent bacteriophage DNA infection. Journal of Molecular Biology. 53(1):159-162.
Michod RM, Wojciechowski M, Hoelzer M. 1988. DNA repair and the evolution of transformation in the bacterium Bacillus subtilis. Genetics (1):118:31-39.
Neumann E, Schaefer-Ridder M, Wang W, Hofschneider PH. 1982. Gene transfer in mouse lyoma cells by electroporation in high electric fields. European Molecular Biology Organization. Journal 1(7):841-845.
Ryu J and Hartin RJ. 1990. Quick transformation in Salmonella typhimurium LT2. Biotechniques 8(1):43-44.
Stanley N. Cohen, Annie C. Y. Chang, and Leslie Hsu. 1972. Nonchromosomal Antibiotic Resistance in Bacteria: Genetic Transformation of Escherichia coli by R-Factor DNA. Proceedings of the National Academy of Sciences of the United States of America. 69(8):2110-2114
Sarkar S, Choudhuri S, Basu T. 2002a. Ethanol-induced enhancement of the transformation of Escherichia.coli by plasmid DNA. Ind. Indian Journal of Biotechnology.1 (2):209-211.
Takahashi R, Valeika SR, Glass KW. 1992. A simple method of plasmid transformation of E.coli by rapid freezing. Biotechniques.13(5):711-715.
Zhiming Tu, Guangyuan He, Kexiu X. Li., Mingjie J. Chen, Junli Chang, Ling Chen, Qing Yao, Dongping P. Liu. 2005. An improved system for competent cell preparation and high efficiency plasmid transformation using different Escherichia coli strains. Electronic Journal of Biotechnology 8(1):114-120.
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