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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...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2012
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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. |
format | Online Article Text |
id | pubmed-3523071 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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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