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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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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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Original Research Articles

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.2018.7(12): 1141-1149
DOI: https://doi.org/10.20546/ijcmas.2018.712.140


A Rapid Method for Titration of Recombinant Baculovirus Stock Based on Bicistronic Expression of Green Fluorescent Protein (GFP) Reporter
Deepika Bisht1, Supriya Bhattacharya1, Lekshmi S. Rajan1, Mashidur Rana2, Ram Bachan2, T. Rama2, Mohini Saini2, and Praveen K. Gupta1*
1Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar-243122, Uttar Pradesh, India
2Division of Biochemistry, Indian Veterinary Research Institute, Izatnagar-243122, Uttar Pradesh, India, India
*Corresponding author
Abstract:

To determine the titer of baculovirus stock accurately based on green fluorescent protein (GFP) reporter, a recombinant baculovirus encoding FLAG-tagged protein and GFP reporter as bicistronic messenger was constructed. The bicistronic messenger RNA was transcribed using polyhedrin promoter and translated in infected Sf21 insect cells. The GFP reporter gene was expressed incap independent manner utilizing internal ribosome entry site (IRES) which helped in titration of baculovirus using end point titration method. The result of this study indicated that recombinant baculovirus expressing GFP reporter as bicistronic messenger facilitated in titration of baculovirus in stock in rapid and accurate manner.


Keywords: Baculovirus, FLAG, Sf21 insect cells, IRES, GFP, PCR
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How to cite this article:

Deepika Bisht, Supriya Bhattacharya, Lekshmi S. Rajan, Mashidur Rana, Ram Bachan, T. Rama, Mohini Saini and Praveen K. Gupta. 2018. A Rapid Method for Titration of Recombinant Baculovirus Stock Based on Bicistronic Expression of Green Fluorescent Protein (GFP) Reporter.Int.J.Curr.Microbiol.App.Sci. 7(12): 1141-1149. doi: https://doi.org/10.20546/ijcmas.2018.712.140