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. 2019 Jun 11;1(8):3015-3022.
doi: 10.1039/c8na00365c. eCollection 2019 Aug 6.

A novel method for genetic transformation of C. albicans using modified-hydroxyapatite nanoparticles as a plasmid DNA vehicle

Ketaki Deshmukh  1 , Sutapa Roy Ramanan  2 , Meenal Kowshik  1
Affiliations

A novel method for genetic transformation of C. albicans using modified-hydroxyapatite nanoparticles as a plasmid DNA vehicle

Ketaki Deshmukh et al. Nanoscale Adv. .

Abstract

In modern biological research, genetic transformation is an important molecular biology technique with extensive applications. In this work, we describe a new method for the delivery of plasmid DNA (pDNA) into a yeast species, Candida albicans. This method is based on the use of novel arginine-glucose-PEG functionalized hydroxyapatite nanoparticles (M-HAp NPs) as a vehicle which delivers pDNA into Candida albicans with a high transformation efficiency of 106 cfu μg-1 of pDNA, without the need for preparation of competent cells. A four-fold higher transformation efficiency as compared to that of the electroporation method was obtained. This new method could provide exciting opportunities for the advancement of the applications of yeasts in the field of biotechnology.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Fig. 1
Fig. 1. (a) XRD pattern, (b) FTIR spectra and (c) TEM micrographs of (i) HAp NPs and (ii) M-HAp NPs. Characteristic XRD peaks of HAp NPs were observed in all the samples. FTIR spectra showed characteristic bands of HAp, amide I, amide II, NH2 glucose and PEG. TEM morphology shows M-HAp NPs with an average length and diameter of 72.7 nm and 8.3 nm.
Fig. 2
Fig. 2. (a) The percentage of bound pDNA quantified using UV absorbance at increasing NP : pDNA ratios. (b) Agarose (0.8%) gel electrophoresis of the M-HApNP–DNA complex with varied NP : pDNA ratios, lane (i) 10 : 1, lane (ii) 50 : 1, lane (iii) 70 : 1, lane (iv) 100 : 1 and lane (v) control pDNA.
Fig. 3
Fig. 3. Transformed C. albicans using M-Hap NPs, without the preparation of competent cells, on kanamycin and geneticin containing SB agar plates (a) and the negative control (b).
Fig. 4
Fig. 4. Agarose gel image of pDNA (i) isolated from C. albicans transformed using M-HAp NPs (ii) and the electroporation technique. (ii) Control pDNA.
Fig. 5
Fig. 5. (a) MTT and (b) resazurin data showing the non-toxic nature of modified HAp NPs on C. albicans after 12, 24 and 36 h of growth at various concentrations ranging from 10–1000 μg mL−1.
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References

    1. Legras J.-L. Merdinoglu D. Cornuet J. Karst F. Mol. Ecol. 2007;16:2091–2102. doi: 10.1111/j.1365-294X.2007.03266.x. - DOI - PubMed
    1. Sicard D. Legras J.-L. C. R. Biol. 2011;334:229–236. doi: 10.1016/j.crvi.2010.12.016. - DOI - PubMed
    1. Botstein D. Fink G. R. Genetics. 2011;189:695–704. doi: 10.1534/genetics.111.130765. - DOI - PMC - PubMed
    1. Khurana V. Lindquist S. Nat. Rev. Neurosci. 2010;11:436–449. doi: 10.1038/nrn2809. - DOI - PubMed
    1. Tenreiro S. Munder M. C. Alberti S. Outeiro T. F. J. Neurochem. 2013;127:438–452. doi: 10.1111/jnc.12271. - DOI - PubMed