Barabensis Opn Cloning Of Differentially Expressed

Rodent is one of the major hidden threat to the agricultural production in China, and Cricetulus barabensis is the dominant rodent species in most Farmland areas at North of China. The osteopontin (OPN) gene is one of the candidate genes about reproductive traits of animals in recent years, which participates almost the entire process of reproduction, and plays an important role in regulating embryo implantation and pregnancy maintenance especially. OPN gene of C. barabensis was cloned and its structure function and the molecular evolution had analyzed, at the same time, expression situation of OPN gene was analyzed for C. barabensis in different developmental stages and different tissues, it is suitable to investigate questions in population outbreak of C.barabensis in the molecular level.In this paper,467bp of OPN gene mRNA including partial exons 2-6, was cloned by using PT-PCR technique in the species of C.barabensis in Shandong Province (accession number in GenBank:HQ647182);its nucleotide sequence, amino acid sequence, protein structure were analyzed and predicted by bioinformatics. The characteristic region in the amino acid sequence——RGD primitives (Arg-Gly-Asp) and highly acidic polyaspartic acid region were found. Besides, a helix occupied the higher proportion in secondary structure of the protein, and hydrophilic was better. The sequence of OPN gene obtained from C. barabensis were compared with the corresponding sequences of Apodemus agrarius, Mus musculus, Rattus norvegicus, Homo sapiens, Bos Taurus, Ovis aries and Sus scrofa, and the phylogentic trees were constructed. It showed that, the relationship between C. barabensis and Apodemus agrarius, Rattus norvegicus, Mus musculus had the closest relationship, but they were distant from Homo sapiens, Bos Taurus and Ovis aries, and had the most distant relationship with Gallus gallus and Coturnix coturnix. The result is consistent with the traditional taxonomic relationship of the above species evolution, which indicates OPN gene is conservative in evolution.The relative expression status of OPN gene in different tissues among different physiological phases of C. barabensis was compared and analysed by the technology of real-time fluorescent quantitative PCR. The results showed that OPN gene was expressed in all the 7 tissues including spleen, kidney, live, heart, small intestine, uterus and ovary, but its expression in different levels were variant, and the relative difference in each physiological phase was also diverse from each other. That may be related to the various biological functions in different tissues of C. barabensis among different physiological phases. According to analyse the expression of OPN gene in uterus and ovary at different developmental stages, it could be indicated that the expression of OPN gene was extremely high in the reproductive organs of C. barabensis during pregnancy and lactation, and it was much higher in pregnancy. The result provides the theoretical evidence at molecular level for demonstrating that the OPN gene plays an important role in pregnancy maintenance, and gives an intuitive proof about the relevance between OPN gene and Animal reproductive performance.