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Hindi Research Article
Chitin Nanowhiskers Mediate Transformation of Escherichia coli by Exogenous Plasmid DNA
Alisa Mera1, Jun Araki2, Takashi Ohtsuki3, Makoto Shimosaka4 and Naoto Yoshida1*1Department of Biochemistry and Applied Biosciences, University of Miyazaki, 1-1 Gakuen Kibanadai-Nishi, Miyazaki 889-2192, Japan
2International Young Researchers Empowerment Center, Shinshu University, Tokida 3-15-1 Ueda, Nagano 386-8567, Japan
3Graduate School of Medicine and Engineering, University of Yamanashi, Kofu 400-8511, Japan
4Division of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, Tokida 3-15-1, Ueda, Nagano 386-8567, Japan
- Corresponding Author:
- Naoto Yoshida, PhD
Department of Biochemistry and Applied Biosciences
University of Miyazaki, 1-1 Gakuen Kibanadai-Nishi
Miyazaki 889-2192, Japan
Tel: +81-985-58-7218
E-mail: a04109u@cc.miyazaki-u.ac.jp
Received date: September 03, 2011; Accepted date: September 26, 2011; Published date: September 28, 2011
Citation: Mera A, Araki J, Ohtsuki T, Shimosaka M, Yoshida N (2011) Chitin Nanowhiskers Mediate Transformation of Escherichia coli by Exogenous Plasmid DNA. J Biotechnol Biomaterial 1:114. doi:10.4172/2155-952X.1000114
Copyright: © 2011 Mera A, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
A colloidal solution of chitin nanowhiskers containing pUC18 plasmid DNA and cells of Escherichia coli was placed on an agar hydrogel and stimulated by sliding friction applied between the agar hydrogel and polystyrene stir stick, leading to transformation of the bacteria to antibiotic resistance. The combination of chitin nanowhiskers and sliding friction was necessary for genetic transformation, indicating that the chitin nanowhiskers induced the Yoshida effect, resulting in the formation of E. coli cells penetration-intermediates. The pUC18 adsorbed onto the chitin nanowhiskers is presumably taken up through the penetration-intermediates, leading to transformation. The transformation efficiency changed when the number of recipient cells and amount of chitin nanowhiskers were varied. The maximum number of penetration-intermediates acquiring pUC18 DNA was obtained with concentrations of agar hydrogel, chitin nanowhiskers, and recipient E. coli cells of 2.5%, 5.0 ?g/ml, and 4.8 x 108 cells/ml, respectively. Optimal transformation efficiency with pUC18 was achieved with 2.1 x 1066 colony forming units/?g of pUC18. Induction of the Yoshida effect with chitin nanowhiskers represents a simpler alternative for introducing genes into bacteria.
