Cloning, expression and phylogenetic analysis of IFN-γ gene in chicken
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Authors
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UTTARA CHATURVEDI
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SHAHINA KALIM
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LAKSHYA VEER SINGH
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A P SAHOO
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S K PALIA
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RAJIV KUMAR
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G RAVI KUMAR
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SANGEETA TIWARI
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ASHOK K TIWARI
Keywords:
Genetic adjuvant, IFN γ, RT-PCRAbstract
In the present study, chicken interferon gamma (Ch.IFN-γ) gene was amplified using specific primers by RT-PCR
and cloned into pTZ 57R/T cloning vector. The clone was confirmed by restriction digestion analysis using KpnI and
ApaI restriction enzymes and orientation was confirmed by colony PCR using T7 promoter primer and gene specific
forward primers. Further, the gene fragment was sub-cloned in eukaryotic expression vector pcDNA 3.1(+) and confirmed
by restriction digestion. The recombinant pcDNA 3.1 having IFN gene (pcDNA.Ch-IFN-γ) was checked for its ability
to express IFN-γ in vitro in cell culture and expression was confirmed by RT-PCR, indicating the gene is functionally
active. The study to evaluate ability of pcDNA.Ch-IFN-γ to combat infection and enhance efficacy of DNA vaccine is
under way.
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