期刊
INTERNATIONAL NANO LETTERS
卷 7, 期 4, 页码 291-295出版社
SPRINGER INTERNATIONAL PUBLISHING AG
DOI: 10.1007/s40089-017-0224-0
关键词
Transformation; Chitosan; pDNA; Nanoparticles; Electrospray
资金
- Tehran University of Medical Sciences & health Services [93-01-87-25230]
A rapid and efficient procedure for DNA transformation is a key prerequisite for successful cloning and genomic studies. While there are efforts to develop a facile method, so far obtained efficiencies for alternative methods have been unsatisfactory (i.e. 10(5)-10(6) CFU/mu g plasmid) compared with conventional method (up to 10(8) CFU/mu g plasmid). In this work, for the first time, we prepared chitosan/pDNA nanoparticles by electrospraying methods to improve transformation process. Electrospray method was used for chitosan/pDNA nanoparticles production to investigate the non-competent bacterial transformation efficiency; besides, the effect of chitosan molecular weight, N/P ratio and nanoparticle size on non-competent bacterial transformation efficiency was evaluated too. The results showed that transformation efficiency increased with decreasing the molecular weight, N/P ratio and nanoparticles size. In addition, transformation efficiency of 1.7 x 10(8) CFU/mu g plasmid was obtained with chitosan molecular weight, N/P ratio and nanoparticles size values of 30 kDa, 1 and 125 nm. Chitosan/pDNA electrosprayed nanoparticles were produced and the effect of molecular weight, N/P and size of nanoparticles on transformation efficiency was evaluated. In total, we present a facile and rapid method for bacterial transformation, which has comparable efficiency with the common method.
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