Volume 4, Issue 5, October 2016, Page: 106-112
Evaluation of the Diphtheria Toxin A Subunit Gene (DT-A) as a Non-conditional Negative Selectable Marker in Tobacco and Rice
Pachamuthu Kannan, Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India
Bharat Bhusan Majhi, Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India
Karuppannan Veluthambi, Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai, Tamil Nadu, India
Received: Jul. 21, 2016;       Accepted: Aug. 1, 2016;       Published: Aug. 29, 2016
DOI: 10.11648/j.jps.20160405.13      View  3464      Downloads  129
Abstract
The diphtheria toxin A subunit gene (DT-A) from Corynebacterium diphtheriae inhibits protein synthesis in eukaryotes. In this study, toxicity of the DT-A gene was evaluated by a transgenic approach in tobacco and rice. The DT-A gene was cloned under transcriptional control of the CaMV 35S promoter and transformed into tobacco. Similarly, CaMV 35S and the maize Ubi1 promoter-driven DT-A gene constructs were transformed into rice. The deployment of the DT-A gene in both tobacco and rice drastically reduced the recovery of transgenic plants in comparison to pCAMBIA1301 (without DT-A). Southern blot analyses of the transgenic plants were done using the hph- and DT-A gene-specific probes to check the presence of the hph and DT-A genes. All the tobacco and rice transgenic plants showed hybridization to junction fragments upon using the hph gene probe. Southern blotting with the DT-A probe revealed that all the transgenic plants either did not have the DT-A gene or harboured truncated DT-A gene in the integrated T-DNAs. None of the transgenic plants carried the complete DT-A gene. The results showed that the DT-A gene can be used as a good non-conditional negative selectable marker in both tobacco and rice. Both CaMV 35S promoter- and Ubi1 promoter-driven DT-A genes were effective as non-conditional negative selectable markers in rice.
Keywords
Agrobacterium tumefaciens, Diphtheria Toxin, Negative Selectable Marker, Rice, Tobacco
To cite this article
Pachamuthu Kannan, Bharat Bhusan Majhi, Karuppannan Veluthambi, Evaluation of the Diphtheria Toxin A Subunit Gene (DT-A) as a Non-conditional Negative Selectable Marker in Tobacco and Rice, Journal of Plant Sciences. Vol. 4, No. 5, 2016, pp. 106-112. doi: 10.11648/j.jps.20160405.13
Copyright
Copyright © 2016 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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