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The 588 bp sequences of two variants of IFNT gene described in this study are novel variants, compared to the variants reported previously in sheep.

In the present study, we have cloned and characterized the IFNT gene of Garole sheep, a popular Indian micro-sheep breed, which is known across the world for its high prolificacy and fecundity. IFNT has been studied extensively in many domesticated and wild ruminant species. Interferon-tau (IFNT), secreted from the elongating blastocyst, acts as the primary signal for maternal recognition of pregnancy in ruminant ungulates. The survivability of embryo, especially during the early embryonic life is dependant on the effective maternal recognition of pregnancy. These findings suggest a potential impact of IFN-T in promoting embryonic growth and development. Here we conclude that the recombinant BuIFN-T was successfully purified to homogeneity from a prokaryotic expression system and it significantly increased the blastocyst production rate in buffalo. Addition of recombinant BuIFN-T (2 μg/ml) significantly improved the rate of blastocyst production, 45.55% against 31.1% control ( p < 0.01). To examine the effect of recombinant BuIFN-T protein on developmental competency of buffalo embryos, purified recombinant BuIFN-T protein was added to in vitro embryo culture medium (at concentration of 0, 1 μg/ml, 2 μg/ml, 4 μg/ml) for 9 days. The purified recombinant BuIFN-T protein was validated by mass spectroscopy analysis. The recombinant BuIFN-T was confirmed by SDS–PAGE and Western blot and subjected to three steps of large scale purification using His Affinity chromatography, Anion Exchange chromatography and Gel Filtration chromatography. After being verified, the fragments without signal sequence, were inserted into the expression vector pET-22b and the recombinant plasmid was induced to express the recombinant protein in a prokaryotic expression system. JX481984, with signal sequence, was obtained through polymerase chain reaction (PCR) from bovine early embryos and was cloned into pJET vector. The buffalo interferon-tau gene (BuIFN-T1) bearing gene bank accession No.

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