A Rapid Method for Titration of Recombinant Baculovirus Stock Based on Bicistronic Expression of Green Fluorescent Protein (GFP) Reporter

Deepika Bisht; Supriya Bhattacharya; Lekshmi S. Rajan; Mashidur Rana; Ram Bachan; T. Rama; Mohini Saini; Praveen K. Gupta

doi:https://doi.org/10.20546/ijcmas.2018.712.140

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Original Research Articles Volume : 7, Issue:12, December, 2018

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Online ISSN : 2319-7706
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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. Gupta^1*

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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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

Copyright: This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license.

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Original Research Articles Volume : 7, Issue:12, December, 2018

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