Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen

Abstract We present a novel method for shockwave-assisted bacterial transformation using a miniature oxyhydrogen detonation-driven shock tube. We have obtained transformation efficiencies of about 1.28 × 106, 1.7 × 106, 5 × 106, 1 × 105, 1 × 105 and 2 × 105 transformants/µg of DNA for Escherichia co...

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Autores principales:	Akshay Datey, Janardhanraj Subburaj, Jagadeesh Gopalan, Dipshikha Chakravortty
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9b3b2984a6564364819cee816d295f76
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spelling	oai:doaj.org-article:9b3b2984a6564364819cee816d295f762021-12-02T15:06:12ZMechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen10.1038/s41598-017-08542-52045-2322https://doaj.org/article/9b3b2984a6564364819cee816d295f762017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08542-5https://doaj.org/toc/2045-2322Abstract We present a novel method for shockwave-assisted bacterial transformation using a miniature oxyhydrogen detonation-driven shock tube. We have obtained transformation efficiencies of about 1.28 × 106, 1.7 × 106, 5 × 106, 1 × 105, 1 × 105 and 2 × 105 transformants/µg of DNA for Escherichia coli, Salmonella Typhimurum, Pseudomonas aeruginosa, Mycobacterium smegmatis, Mycobacterium tuberculosis (Mtb) and Helicobacter pylori respectively using this method which are significantly higher than those obtained using conventional methods. Mtb is the most difficult bacteria to be transformed and hence their genetic modification is hampered due to their poor transformation efficiency. Experimental results show that longer steady time duration of the shockwave results in higher transformation efficiencies. Measurements of Young’s modulus and rigidity of cell wall give a good understanding of the transformation mechanism and these results have been validated computationally. We describe the development of a novel shockwave device for efficient bacterial transformation in complex bacteria along with experimental evidence for understanding the transformation mechanism.Akshay DateyJanardhanraj SubburajJagadeesh GopalanDipshikha ChakravorttyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Akshay Datey Janardhanraj Subburaj Jagadeesh Gopalan Dipshikha Chakravortty Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
description	Abstract We present a novel method for shockwave-assisted bacterial transformation using a miniature oxyhydrogen detonation-driven shock tube. We have obtained transformation efficiencies of about 1.28 × 106, 1.7 × 106, 5 × 106, 1 × 105, 1 × 105 and 2 × 105 transformants/µg of DNA for Escherichia coli, Salmonella Typhimurum, Pseudomonas aeruginosa, Mycobacterium smegmatis, Mycobacterium tuberculosis (Mtb) and Helicobacter pylori respectively using this method which are significantly higher than those obtained using conventional methods. Mtb is the most difficult bacteria to be transformed and hence their genetic modification is hampered due to their poor transformation efficiency. Experimental results show that longer steady time duration of the shockwave results in higher transformation efficiencies. Measurements of Young’s modulus and rigidity of cell wall give a good understanding of the transformation mechanism and these results have been validated computationally. We describe the development of a novel shockwave device for efficient bacterial transformation in complex bacteria along with experimental evidence for understanding the transformation mechanism.
format	article
author	Akshay Datey Janardhanraj Subburaj Jagadeesh Gopalan Dipshikha Chakravortty
author_facet	Akshay Datey Janardhanraj Subburaj Jagadeesh Gopalan Dipshikha Chakravortty
author_sort	Akshay Datey
title	Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
title_short	Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
title_full	Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
title_fullStr	Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
title_full_unstemmed	Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
title_sort	mechanism of transformation in mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/9b3b2984a6564364819cee816d295f76
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Mechanism of transformation in Mycobacteria using a novel shockwave assisted technique driven by in-situ generated oxyhydrogen

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