Please use this identifier to cite or link to this item: http://nopr.niscair.res.in/handle/123456789/43344
Title: Expression of Omp16 and L7/L12 Brucella abortus protective antigens as secretory fusion proteins in mammalian cells
Authors: Prusty, Bikash R
Tabassum, Rizwana
Chaudhuri, Pallab
Saini, Mohini
Chaturvedi, V K
Mishra, B P
Gupta, Praveen K
Keywords: Brucella abortus;Omp16;L7/L12;Fusion gene;Codon optimization;Heterologous gene expression
Issue Date: Jul-2017
Publisher: NISCAIR-CSIR, India
Abstract: Brucella abortus is a facultative intracellular bacterial pathogen infecting animals and humans. This preliminary study was designed to express two immunogenic genes of Brucella abortus as recombinant fusion proteins in mammalian cells for being studied as a vaccine candidate in mammalian hosts, especially mice and cattle, in our next study. The complete open reading frame sequences of two immune dominant genes of Brucella abortus namely, Omp16 and L7/L12 were fused with an intermediate spacer along with N-terminal fusion with secretory signal sequence from immunoglobulin M. The complete fusion gene sequence was codon optimized for expression in mammalian cells. For expression analysis, the codon optimized synthetic gene was cloned in pDsRed-Express-N1 mammalian expression vector, with C-terminal fusion with red fluorescent protein sequence. On transfection in MDBK and HEK-293 cells, appearance of red fluorescence in transfected cells indicated expression of Omp16- L7/L12 fusion proteins along with RFP. The Omp16-L7/L12 fusion construct without RFP sequence was also expressed in mammalian cells. The expressed Omp16-L7/L12 fusion proteins were confirmed through both indirect fluorescence antibody test and western blot. This preliminary study suggested that the codon optimized Omp16-L7/L12 fusion construct is ready to be studied in hosts like mice and cattle for its vaccine efficacy.
Page(s): 289-295
URI: http://nopr.niscair.res.in/handle/123456789/43344
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.16(3) [July 2017]

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