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Functional verification of the diphtheria toxin A gene in a recombinant system

Diphtheria toxin A (DTA), a segment of the diphtheria toxin (tox), inhibits protein synthesis in cells. When released from a cell, DTA is nontoxic and cannot enter other cells independently without the help of diphtheria toxin B. In this study, we artificially synthesized the DTA gene sequence and c...

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Autores principales: Zhang, Jingfeng, Wei, Hengxi, Guo, Xinzheng, Hu, Minghua, Gao, Fenglei, Li, Li, Zhang, Shouquan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3523071/
https://www.ncbi.nlm.nih.gov/pubmed/23062032
http://dx.doi.org/10.1186/2049-1891-3-29
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author Zhang, Jingfeng
Wei, Hengxi
Guo, Xinzheng
Hu, Minghua
Gao, Fenglei
Li, Li
Zhang, Shouquan
author_facet Zhang, Jingfeng
Wei, Hengxi
Guo, Xinzheng
Hu, Minghua
Gao, Fenglei
Li, Li
Zhang, Shouquan
author_sort Zhang, Jingfeng
collection PubMed
description Diphtheria toxin A (DTA), a segment of the diphtheria toxin (tox), inhibits protein synthesis in cells. When released from a cell, DTA is nontoxic and cannot enter other cells independently without the help of diphtheria toxin B. In this study, we artificially synthesized the DTA gene sequence and cloned it into pEGFP-N1 to generate the recombinant vector pEGFP-N1-DTA. This recombinant vector was then transfected into 293T cells to observe the effect of DTA protein expression on enhanced green fluorescent protein (EGFP) protein expression and the proliferation of 293T cells. After 48 h, high levels of EGFP expression were seen in control pEGFP-N1-transfected cells, whereas very low levels were seen in cells transfected with pEGFP-N1-DTA. Reverse transcription polymerase chain reaction confirmed the expression of the DTA gene in cells transfected with pEGFP-N1-DTA. Further, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed a significant difference in cell proliferation between the control group and the pEGFP-N1-DTA-transfected group. Using the expression of EGFP expression as an indicator, this study revealed that DTA expression can inhibit intracellular protein synthesis and cell proliferation.
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spelling pubmed-35230712012-12-16 Functional verification of the diphtheria toxin A gene in a recombinant system Zhang, Jingfeng Wei, Hengxi Guo, Xinzheng Hu, Minghua Gao, Fenglei Li, Li Zhang, Shouquan J Anim Sci Biotechnol Research Diphtheria toxin A (DTA), a segment of the diphtheria toxin (tox), inhibits protein synthesis in cells. When released from a cell, DTA is nontoxic and cannot enter other cells independently without the help of diphtheria toxin B. In this study, we artificially synthesized the DTA gene sequence and cloned it into pEGFP-N1 to generate the recombinant vector pEGFP-N1-DTA. This recombinant vector was then transfected into 293T cells to observe the effect of DTA protein expression on enhanced green fluorescent protein (EGFP) protein expression and the proliferation of 293T cells. After 48 h, high levels of EGFP expression were seen in control pEGFP-N1-transfected cells, whereas very low levels were seen in cells transfected with pEGFP-N1-DTA. Reverse transcription polymerase chain reaction confirmed the expression of the DTA gene in cells transfected with pEGFP-N1-DTA. Further, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed a significant difference in cell proliferation between the control group and the pEGFP-N1-DTA-transfected group. Using the expression of EGFP expression as an indicator, this study revealed that DTA expression can inhibit intracellular protein synthesis and cell proliferation. BioMed Central 2012-10-15 /pmc/articles/PMC3523071/ /pubmed/23062032 http://dx.doi.org/10.1186/2049-1891-3-29 Text en Copyright ©2012 Zhang et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Zhang, Jingfeng
Wei, Hengxi
Guo, Xinzheng
Hu, Minghua
Gao, Fenglei
Li, Li
Zhang, Shouquan
Functional verification of the diphtheria toxin A gene in a recombinant system
title Functional verification of the diphtheria toxin A gene in a recombinant system
title_full Functional verification of the diphtheria toxin A gene in a recombinant system
title_fullStr Functional verification of the diphtheria toxin A gene in a recombinant system
title_full_unstemmed Functional verification of the diphtheria toxin A gene in a recombinant system
title_short Functional verification of the diphtheria toxin A gene in a recombinant system
title_sort functional verification of the diphtheria toxin a gene in a recombinant system
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3523071/
https://www.ncbi.nlm.nih.gov/pubmed/23062032
http://dx.doi.org/10.1186/2049-1891-3-29
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