Abstract:
Cyclin dependent kinase inhibitors B (CDKNIB, namely p27) inhibits cyclin-cyclin dependent kinase complexes and halts cell cycle progression.After cloning p27 gene sequence of dairy goat and elucidating its biological characteristics, RNA interference and gene overexpression techniques were used to explore the function of p27 gene on the proliferation of mammary epithelial cells. Taken mammary tissue from Xinong Saanen dairy goats as experimental material, the whole CDS sequence of p27 gene was cloned and the bioinformatics analysis of p27 gene coding sequence was conducted. Results showed that the complete CDS region of p27 gene was 597 bp, encoding 197 amino acids, and p27 protein was a hydrophilic unstable protein without transmembrane spiral structure. The expression of p27 gene in mammary tissue of different stages was measured by RT-PCR, and higher expression level was detected at lactation stage compared with that at dry period. The eukaryotic expression plasmid of p27 gene was transfected into mammary epithelial cells, and its effects on the proliferation of mammary epithelial cells were evaluated by CCK-8, Edu staining and Western blot. Results showed that overexpression of p27 gene inhibited the proliferation and DNA synthesis of mammary epithelial cells, and increased the expression of CDK2(P
