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The present study we have undertaken is to clone and express the partial length cDNA of extra embryonic tissue-spermatogenesis homeobox gene 1 (ESX-1). ESX-1 is an X-linked gene which encodes for a homeobox protein. The ESX-1 expressed specifically in extraembryonic tissues during development and in the adult testis as well. In particular, in situ hybridization experiments delimited ESX-1 transcription to preleptotene spermatocytes and round spermatids, suggesting a
role in spermatogenesis. Available Bos taurus ESX-1 gene sequence was used to design the gene specific primers and used for amplification of Bubalis bubalus ESX-1 gene from the cDNA. Amino acid sequence has shown his hest identity with Bos taurus (91.6%). The homeobox region has shown the identity of 98.4%, 70.5%, 70.5%, 69.4% and 69.4% with Bos taurus, Homo sapiens,
Pan troglodytes, Macaca mulatta and Mus musculus respectively. The ESX-1 cDNA was cloned and expressed in Escherichia coli and was purified in a single chromatographic step using Cobalt-Agarose gel affinity column. The purity and the molecular weight of the fusion protein were checked in SDS-PAGE which showed a purified product of molecular weight 27KDa. The purified
product was confirmed by Western blot analysis using Ni- HRPase conjugate. As the ESX-1 protein is X-chromosome specific and expressed in only X-bearing spermatozoa the antibodies raised against the protein can be used for assessing the enrichment of buffalo spermatozoa in sex sorted semen.
N Vijay has completed his BVSc & AH degree from Veterinary College, Bangalore in the year 2008. He got selected in Indian Veterinary research
Institute, Izatnagar to pursue Masters Degree in the discipline Animal Biochemistry in the year 2009 and completed the same with MVSc degree in
2011. Presently he is pursuing PhD from Deemed University, Indian Veterinary Research Institute, Izatnagar.
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