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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): 1656-1668
DOI: https://doi.org/10.20546/ijcmas.2018.712.193


Synthesis and Evaluation of Gyps vulture Multidrug Resistant Protein 4 (MRP4/ABCC4) Specific Multiple Antigenic Peptide (MAP) for its Immunoreactivity
Mashidur Rana1, Mohini Saini1*, Sasmita Barik1, Ram Bachan1, Deepika Bisht2, Supriya Bhattacharya2, Kirthika P.3, Praveen Singh3, Satish Kumar3 and Praveen K. Gupta2
1Division of Biochemistry
2Division of Veterinary Biotechnology
3Central Instrumentation and Bioengineering Facility, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, U.P-243122, India
*Corresponding author
Abstract:

Efforts are in progress to conserve critically endangered Gyps spp. vultures by identifying and removing the vulture-unsafe drugs from the food chain. Several drugs namely diclofenac, aceclofenac, ketoprofen and nimesulide are found to cause hyperuricemia and fatal nephrotoxicity in Gyps vultures. Research on development of an in vitro model for screening drugs in practice and the new molecules for their hyperuricemic potential can facilitate the conservation efforts. Multi drug resistant protein 4 (MRP4/ABCC4) is an efflux transporter on the luminal side of proximal renal tubular membrane that plays key role in urate as well as drug transport. In the present study, we report development and use of Gyps himalayensis MRP4 specific multiple antigenic peptide (MAP). Sequences of linear peptide and N terminal immunodominant region of Gyps himalayensis MRP4 was predicited in silico. Tetrameric MAP was synthesized in solid phase synthesis (SPPS) using Wang resin and employed for immunization of rabbit for obtaining polyclonal serum. N terminal region (1-737 amino acids) of vulture MRP4 predicted to contain immunodominant epitopes was expressed in E. coli BL21 cells and purified in Ni-NTA affinity chromatography. The 24 kDa purified protein could be detected in immunoblots using anti-MAP serum indicating its potential in detection of MRP4 expression. This hyperimmune serum can be exploited for its use in detection of MRP4 expression in avian renal tubular cells or MRP4 expressing cells before their use for screening drugs for their role in urate transport.


Keywords: Gyps himalayensis, Vulture, MRP4 (Multi Drug Receptor Protein 4), ABC (ATP binding cassette) superfamily, MAP (Multi Antigenic Peptide), SPPS (Solid Phase Peptide Synthesis)
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How to cite this article:

Mashidur Rana, Mohini Saini, Sasmita Barik, Ram Bachan, Deepika Bisht, Supriya Bhattacharya, P. Kirthika, Praveen Singh, Satish Kumar and Praveen K. Gupta. 2018. Synthesis and Evaluation of Gyps vulture Multidrug Resistant Protein 4 (MRP4/ABCC4) Specific Multiple Antigenic Peptide (MAP) for its Immunoreactivity.Int.J.Curr.Microbiol.App.Sci. 7(12): 1656-1668. doi: https://doi.org/10.20546/ijcmas.2018.712.193