In order to analyze the expression of canine p27 protein in canine mammary gland tumors,the recombinant plasmid pET-p27 was constructed for the prokaryotic expression.New Zealand white rabbits were immunized with the purified recombinantprotein p27 to prepare the anti-canine p27 polyclonal antibody,and the expression of canine p27 protein in 84 cases of canine breast tumor was analyzed using this antibody.The results showed that the antibody titer was more than 1∶12,800 and had a specific response to the recombinant protein based on western blot analysis.Immunohistochemistry assay demonstrated that the expression rate of canine p27 protein in canine malignant mammary gland tumor was lower than that in benign tumor,which indicated that the level of canine p27 expression might be related to the degree of canine mammary gland tumor malignant.Our research could provide basis for the further study of the biological function of canine p27 protein.%为分析犬p27蛋白在犬乳腺肿瘤组织中的表达情况,本研究构建了原核表达重组质粒pET-p27,进行重组蛋白的表达,并将纯化的重组蛋白免疫新西兰白兔,制备了抗犬p27蛋白多克隆抗体.将制备的犬p27抗体对临床84例犬乳腺肿瘤组织中的犬p27蛋白表达情况进行了检测.结果显示,制备的抗体效价达1∶12 800以上;western blot检测显示,该多克隆抗体能够与重组蛋白发生特异性反应;免疫组织化学试验表明,犬p27蛋白在犬恶性乳腺肿瘤组织中表达率低于犬良性乳腺肿瘤组织,提示犬p27的表达水平可能与犬乳腺肿瘤的恶性程度相关,本实验为进一步研究犬p27蛋白的生物学功能奠定了基础.
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