Artificial transformation methodologies for improving the efficiency of plasmid DNA transformation and simplifying its use

Ren, Jun; Karna, Sandeep; Lee, Hyang-Mi; Yoo, Seung Min; Na, Dokyun

doi:10.1007/s00253-019-10173-x

Cited by 12 publications

(13 citation statements)

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“…In bacteria, CPPs have been used without additional treatments, but recently, the chemical treatment of cells has improved the delivery efficiency of CPP conjugates 46 . Electroporation has been widely used as an effective method for biomacromolecule delivery, and electroporation is generally more efficient than chemical treatments, such as that with CaCl 2 51 . In this study, we developed and optimized a new electroporationbased delivery method to further improve CPP delivery efficiency, and used the method for CPP delivery experiments.…”

Section: Resultsmentioning

confidence: 99%

Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

et al. 2021

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In bacterial biotechnology, instead of producing functional proteins from plasmids, it is often necessary to deliver functional proteins directly into live cells for genetic manipulation or physiological modification. We constructed a library of cell-penetrating peptides (CPPs) capable of delivering protein cargo into bacteria and developed an efficient delivery method for CPP-conjugated proteins. We screened the library for highly efficient CPPs with no significant cytotoxicity in Escherichia coli and developed a model for predicting the penetration efficiency of a query peptide, enabling the design of new and efficient CPPs. As a proof-of-concept, we used the CPPs for plasmid curing in E. coli and marker gene excision in Methylomonas sp. DH-1. In summary, we demonstrated the utility of CPPs in bacterial engineering. The use of CPPs would facilitate bacterial biotechnology such as genetic engineering, synthetic biology, metabolic engineering, and physiology studies.

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Section: Resultsmentioning

confidence: 99%

Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

et al. 2021

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“…The development of competent cells is one of the key techniques for introducing foreign DNA into cells. Many literature have reported the development of chemotransformation, physical-assisted transformation, electrotransformation and other new nanocarrier-mediated transformation (Chan et al, 2013;Choi et al, 2013;Brito et al, 2017;Roychoudhury, Basu & Sengupta, 2009;Ren et al, 2019). Among these methods, the chemical artificial transformation was regarded as the simple and efficient bacterial transformation technique.…”

Section: Discussionmentioning

confidence: 99%

“…The ideal parameters to obtain maximum transformation efficiency in E. coli DH5α strain with pUC19 plasmid DNA was Sr 2+ of 90 mM, heat-shock time of 90 s, the amount of pUC19 plasmid DNA of 9 ng. However, the precise details of transformation mechanisms remain vague (Aich et al, 2012;Claverys & Martin, 2003;Wang, Sun & Ma, 2016;Ren et al, 2019). Some researchers considered that cell membranes fluidity was weakened by using cations (e.g., Na + , Mg 2+ , Ca 2+ , Ba 2+ ) and low temperature (ice-water bath).…”

Section: Discussionmentioning

confidence: 99%

“…Thereafter, the chemical transformation methods (Nørgard, Keem & Monahan, 1978;Dagert & Ehrlich, 1979;Zhang, Xu & Xu, 2004) have continuously been modified with various combinations of chemical solutions including different cations (Mg 2+ , Ca 2+ , Sr 2+ , Ba 2+ , Mn 2+ , K + , Na + , Rb + ), PEG, DMSO (Chung & Miller, 1988;Chan et al, 2013) and glycerin Shanehbandi et al (2013). Considering the practicality and convenience of different artificial methods, more efficient exogenous gene transfer systems also have been developed by physical or chemical methods, including chemical procedures (Inoue, Nojima & Okayama, 1990;Sarkar, Choudhuri & Basu, 2002a;Sarkar, Choudhuri & Basu, 2002b;Song et al, 2007), high-voltage electroporation (Dower, Miller & Ragsdale, 1988;Dunny, Lee & LeBlanc, 1991;Sheng, Mancino & Birren, 1995), a biolistic propulsion system (Shark et al, 1991), liposome-mediated DNA transfer (Kawata, Yano & Kojima, 2003), microwaves and ultrasounds assisted DNA transfer (Zarnitsyn & Prausnitz, 2004;Fregel, Rodriguez & Cabrera, 2008;Tripp, Maza & Young, 2013;Deeks et al, 2014) and chemicalphysical (Rb + , sepiolite and nanomaterials) induced transformation (Ren et al, 2017;Ren, Na & Yoo, 2018;Ren et al, 2019), etc. The transformation efficiency (10 4∼10 8 CFU/µg DNA) of different strains can obviously be obtained from these methods described.…”

Section: Introductionmentioning

confidence: 99%

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Effects of Sr^2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation

Wang

et al. 2020

PeerJ

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Bacterial gene transformation used with Escherichia coli as a desired microorganism is one of the important techniques in genetic engineering. In this study, the preparation of E. coli DH5α competent cells treated with SrCl2 and transformation by heat-shock with pUC19 plasmid was optimized by Response Surface Methodology (RSM). Other five E. coli strains including BL21 (DE3), HB-101, JM109, TOP10 and TG1, three different sizes plasmids (pUC19, pET32a, pPIC9k) were used to verify the protocol, respectively. The transformation mechanism was explored by scanning electron microscope combined with energy dispersive spectrometer (SEM-EDS), atomic absorption spectroscopy (AAS) and Fourier-transform infrared spectroscopy (FT-IR). An equation of regression model was obtained, and the ideal parameters were Sr2 + ions of 90 mM, heat-shock time of 90 s and 9 ng of plasmid. Under this conditions, the transformation efficiency could almost reach to 106 CFU/µg DNA. A small change of the cell surface structure has been observed between E. coli DH5α strain and competent cells by abovementioned spectrum technologies, which implied that a strict regulation mechanism involved in the formation of competent cells and transformation of plasmids. An equation of regression model for the competent cells preparation and plasmid transformation could be applied in gene cloning technology

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“…Furthermore, even in species and cultivars where plant transformation is possible, the process is expensive, slow, requires significant resources in terms of facilities and expertise, is frequently inefficient, and damages the plant genome. (Doyle et al, 2019) Various and equally vexing challenges of the type referred to by Doyle et al (2019) target organisms-bacteria, fungi, plants, and animalswith each requiring customized methods with many methods working for only some species of each group (e.g., Yoshida and Sato, 2009;Rivera et al, 2014;Kelliher et al, 2019;Ren et al, 2019;Lule-Chávez et al, 2020). The difficulty in developing and deploying customized methods and materials, along with highly specialized expertise, limits the ability of gene technology to amplify the risks.…”

Section: Species Rangementioning

confidence: 99%

Differentiated impacts of human interventions on nature

Heinemann

Paull

Walker

et al. 2021

Elementa: Science of the Anthropocene

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Biotechnology describes a range of human activities in medicine, agriculture, and environmental management. One biotechnology in particular, gene technology, continues to evolve both in capacity and potential to benefit and harm society. The purpose of this article is to offer a policy bridge from unproductive descriptions of gene technology to useful methods for identifying sources of significant biological and socioeconomic risk in complex food systems. Farmers and the public could be voluntarily and involuntarily interacting with new techniques of genome editing and gene silencing in entirely new ways, limiting the usefulness of previous gene technology histories to predict safety. What we believe is a more consistent, verifiable, and practical approach is to identify the critical control points that emerge where the scale effects of a human activity diverge between risk and safety. These critical control points are where technical experts can collaborate with publics with different expertise to identify and manage the technology. The use of technical terminology describing biochemical-level phenomena discourages publics that are not technical experts from contesting the embedded cultural perspectives and uncertainty in “scientific” concepts and prejudice the risk discourse by ignoring other issues of significance to society. From our perspective as gene technologists, we confront the use of pseudo-scale language in risk discourse and propose an escape path from clashes over whether risks that arise spontaneously (from nature) can be perfectly mimicked by gene technology to a discussion on how to best control the risks created by human activity. Scale is conceptually implicit and explicit in gene technology regulation, but there is no agreement about what scales are most useful to managing risk and social expectations. Both differentiated governance (risk-tiered) and responsible research and innovation models could accommodate the critical control points mechanism that we describe.

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Artificial transformation methodologies for improving the efficiency of plasmid DNA transformation and simplifying its use

Cited by 12 publications

References 64 publications

Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

Effects of Sr^2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation

Differentiated impacts of human interventions on nature

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Artificial transformation methodologies for improving the efficiency of plasmid DNA transformation and simplifying its use

Cited by 12 publications

References 64 publications

Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

Effects of Sr2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation

Differentiated impacts of human interventions on nature

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Product

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Effects of Sr^2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation