奶牛子宫内膜粘液抗菌肽的分离纯化

Isolation and Purification of Antibacterial Polypeptide from Cow Uterus Mucus

为研发奶牛抗菌肽生物制剂,防治奶牛子宫内膜炎,本研究采用酸性尿素聚丙烯酰胺凝胶电泳琼脂糖弥散法检测子宫内膜粘液酸溶性提取物抗菌活性;应用琼脂糖电泳及水平电泳洗脱分离相应的抗菌条带,HPLC进一步分离纯化,琼脂糖弥散法检测纯化分子抗菌活性,Tricine-SDS-PAGE电泳检测其分子量。结果表明,从奶牛子宫内膜酸溶性提取物中纯化出一分子量约为14kD左右的抗菌多肽,对大肠杆菌(E.coliBL21)和金黄色葡萄球菌26003具有抗菌活性。本研究可能分离纯化出一种新的子宫内膜分泌的抗菌多肽,其可能参与奶牛子宫的天然防御机制。

To research and development biological agents of Dairy Cow antimicrobial peptides, prevent and treat endometritis cow disease, acid-soluble extract of cow uterus mucus was obtained and antibacterial activity was detected by gel overlay technique, then it was further isolated and purified by HPLC and analyzed antibacterial activity by agarose radial diffusion assay. The molecular weight was determined by Trieine-SDS-PAGE. The results showed that the antibacterial polypeptide was able to effectively kill E. coli BL21 and the Staphylococcus bacteria 26003, its molecular weight was about 14kDa. It may be a new antibacterial polypeptide from cow uterus mucus and take part in natural immunology defense in uterus mucus.