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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 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2017: 100.00
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2017: 100.00
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 2017: 100.00
NAAS RATING 2018: 5.38

Int.J.Curr.Microbiol.App.Sci.2019.8(6): 2828-2836
DOI: https://doi.org/10.20546/ijcmas.2019.806.341


Cloning, Expression and in silico Characterization of a Truncated Antiviral Protein Gene from Bougainvillea spectabilis Willd.
Magar Nakul Divakar1, S. Rajesh1, P. Renukadevi2 and B. Rajagopal1,3*
1Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore- 641003, India
2Department of Sericulture, FC&RI, Mettupalayam- 641301, India
3Department of Fruit Science, HC&RI, Periyakulam-625601, India
*Corresponding author
Abstract:

Bougainvillea antiviral protein (BAP) is one among a class of the ribosomal inactivating proteins isolated from Bougainvillea spectabilis willd. Truncated version of the BAP gene was cloned and expressed in a prokaryotic vector to abolish its cytotoxicity. RNA was isolated from mature leaves of Bougainvillea and the full length cDNA was amplified by reverse transcription-PCR using template mRNA. This full length cDNA of size 756 bp was amplified using the proofreading polymerase (Q5 polymerase) and end to end gene specific primers for removal of C-terminal, the amplicon was cloned in pJET1.2l vector by blunt end cloning method. Restriction digestion was performed to release the fragment which was further ligated into prokaryotic expression vector pET29a. The recombinant plasmid was transferred into E.coli expression strain BL21 (DE3) and the truncated-BAP gene was expressed by isopropyl β-D thiogalactopyranoside (IPTG) induction. Transformed colonies expressed recombinant fusion Bougainvillea antiviral protein of molecular weight ~14.6 kDa, the size expected for the truncated BAP gene.


Keywords: Bougainvillea spectabilis Willd., Q5 Polymerase, IPTG
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How to cite this article:

Magar Nakul Divakar, S. Rajesh, P. Renukadevi and Rajagopal, B. 2019. Cloning, Expression and in silico Characterization of a Truncated Antiviral Protein Gene from Bougainvillea spectabilis Willd..Int.J.Curr.Microbiol.App.Sci. 8(6): 2828-2836. doi: https://doi.org/10.20546/ijcmas.2019.806.341