Abstract：Abstract The aim of the study was to clone the CDS of retinol binding protein 4 gene (RBP4), and to determine its expression in different months old testis of sheep (Ovis aires), which might provide basis for its functional research on testis development. Sheep testis was used as material in this study. The sequence of RBP4 gene was cloned by RT-PCR, and structure of RBP4 protein was analyzed with relevant bioinformatics softwares. The mRNA and protein expression of RBP4 was determined using qRT-PCR and immunohistochemistry, respectively. The results showed that the CDS length of RBP4 gene was 606 bp, and it encoded goat (Capra hircus), cow (Bos taurus), pig (Sus scrofa), human (Homo sapiens) and mouse (Mus musculus), respectively. The sheep RBP4 gene was the closest with that of goat, followed by cow, pig, human and mouse. RBP4 mRNA expressed in all different months old and the expression was firstly decreased and then increased with month increasing. The difference between the adjacent two groups were extremely significant (P＜0.01). RBP4 was mainly localized in the primary spermatocytes, spermatid, spermatozoon and seminiferous tubule. The positive reaction integrated optical density showed a trend of first increasing and then decreasing with month increasing, which was opposite to the expression of RBP4 mRNA. The study result indicated that the RBP4 gene may participate in the control of sheep testis development, which provides reference to further exploration of the influence of RBP4 in sheep reproductive process.
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