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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2020)
[Effective from January 1, 2020]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2018 - IJCMAS--ICV 2018: 95.39
For more details click here

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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706
Issues : 12 per year
Publisher : Excellent Publishers
Email : editorijcmas@gmail.com / submit@ijcmas.com
Editor-in-chief: Dr.M.Prakash
Index Copernicus ICV 2018: 95.39
NAAS RATING 2020: 5.38

Int.J.Curr.Microbiol.App.Sci.2017.6(9): 1718-1724
DOI: https://doi.org/10.20546/ijcmas.2017.609.212


Isolation, Cloning and Expression of Insecticidal-Protein-Encoding Gene tcdA from Photorhabdus luminescens in Escherichia coli
S.R. Mulla, Shourabh Joshi* and Joy Das
1Department of Biotechnology and Crop improvement, University of Horticultural Science, Bagalkot-587103, India
2Department of Biotechnology, University of Agricultural Sciences, GKVK, Bangalore -560065, Karnataka, India
*Corresponding author
Abstract:

In this study, the tcdA coding gene of Photorhabdus luminescens was inserted in pET32a (+) plasmid with extra His-tag sequence. Photorhabdus luminescens strain was isolated from Western Ghats of Karnataka, which is symbiotically associated with entomopathogenic nematode Heterorhabditis spp. The DNA sequence of cloned toxin gene (7551 bp) has an open reading frame encoding 2506 amino acids with a predicted molecular mass of 283kDa. The integrity of the constructed plasmid was confirmed using restriction digestion and PCR. The tcdA gene was expressed after induction with IPTG in Escherichia coli BL21. Recombinant tcdA was purified by Ni- NTA Affinity Chromatography.


Keywords: tcdA, Photorhabdus luminescens, Cloning and expression.
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How to cite this article:

Mulla, S.R., Shourabh Joshi and Joy Das. 2017. Isolation, Cloning and Expression of Insecticidal-Protein-Encoding Gene tcdA from Photorhabdus luminescens in Escherichia coli.Int.J.Curr.Microbiol.App.Sci. 6(9): 1718-1724. doi: https://doi.org/10.20546/ijcmas.2017.609.212