Molecular characterization and expression profile of interferon-stimulated gene 15 (ISG15) in the endometrium of goat (Capra hircus)

Abstract

Interferon-stimulated gene 15 (ISG15), a ubiquitin-like protein, is responsible for uterine receptivity, implantation and conceptus development in different ruminant species, but in goat (Capra hircus) its role is yet to be explicated. In the present study, the ISG15 gene was cloned, characterized and its temporal expression profile was examined in the endometrium of caprine (cp). A fragment of cpISG15 gene, 1033 bp in length, was amplified, cloned and sequenced from genomic DNA covering the coding region. Sequence analysis of cpISG15 gene revealed that it was comprised of two exons of 59 bp and 496 bp encoding a peptide of 157 amino acids. Complementary DNA (cDNA) and deduced amino acid sequences of cpISG15 exhibited 99 and 98, 93 and 88, 94 and 89, 76 and 66, and 73 and 62% identity with that of sheep, cattle, buffalo, human and mice, respectively. Further, relative expression of cpISG15 mRNA and protein was determined by quantitative real-time PCR (qPCR) and Western blot, respectively, in the endometrium of pregnant and cyclic does. Both cpISG15 mRNA and protein were expressed maximally (P < 0.05) in the endometrium during early stage of pregnancy (16–24 d) as compared to cyclic does, but no significant difference was observed in cpISG15 mRNA and protein expression in the endometrium between the later stage of pregnancy (25–40 d) and cyclic does. It is concluded that cpISG15 is almost similar in structure and probably in function also to other species as it has been found significantly upregulated during early pregnancy.