Functional verification of the diphtheria toxin A gene in a recombinant systemReportar como inadecuado




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Journal of Animal Science and Biotechnology

, 3:29

Animal Genetics

Abstract

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.

KeywordsDiptheria toxin A Cell proliferation Protein synthesis Electronic supplementary materialThe online version of this article doi:10.1186-2049-1891-3-29 contains supplementary material, which is available to authorized users.

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Autor: Jingfeng Zhang - Hengxi Wei - Xinzheng Guo - Minghua Hu - Fenglei Gao - Li Li - Shouquan Zhang

Fuente: https://link.springer.com/







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