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|Title:||Sequencing of Mx gene in an indian major carp|
|Agrotags:||Rohu, Interferon, Poly I: C, PBS, Mx, gene, dsRNA, Virus, RT-PCR, DNA, sequence.|
|Abstract:||Mx gene is inducible by type I interferon and is involved in antiviral defenses. Interferons are pleiotropic glycoproteins belonging to a large family of Cytokines. The type I interferon constitute the first line of defence against viral infection in the vertebrate innate immune system. Mx gene inducible by type I interferon, natural and synthetic double stranded RNA (ds RNA) and unmethylated bacterial CpG motif, both in fish and mammalian hosts. Mx expression was observed in fish after viral infection (i.e. IPNV, VHSV, ISAV), although the antiviral role of Mx proteins in fish remains unclear. Mx gene has been cloned and sequenced in many fishes including gold fish and grass carp of family Cyprinidae. The aim of the present study was to standardize the RT-PCR protocol for the expression of Mx gene in an Indian major carp, rohu (Labeo rohita). Mx response was measured in rohu injected with the synthetic double stranded polyribonucleotide Poly I: C (500 μg/fish) and the control fish was injected with PBS alone. Total RNA was extracted from liver of acclimatized fish after 3 days of post-injection. It was transcribed to cDNA and amplification of cDNA was done by RT-PCR. The sequence was obtained from purified gel product. The blasts search was done for confirmation of Mx gene in rohu. It was found 99% matching with grass carp Mx and 98% matching with gold fish Mx3 belonging to family Cyprinidae. These results confirm the presence of Mx gene expression in an Indian major carp, rohu. The obtained partial sequence can be used for taxonomic and phylogenetic purposes.|
|Appears in Collections:||M. Sc. Dissertations|
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