| 奶牛子宫内膜组织抗菌肽分离纯化及其抗菌活性研究 |
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| 引用本文： | 王东升,;米宝明,;荔霞,;张世栋,;李世宏,;宗瑞谦,;祝秉东,;王桂荣,;严作廷.奶牛子宫内膜组织抗菌肽分离纯化及其抗菌活性研究[J].中兽医医药杂志,2014(6):24-27. |
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| 作者姓名： | 王东升 ;米宝明 ;荔霞 ;张世栋 ;李世宏 ;宗瑞谦 ;祝秉东 ;王桂荣 ;严作廷 |
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| 作者单位： | [1]中国农业科学院兰州畜牧与兽药研究所中国农业科学院临床兽医学研究中心,甘肃兰州730050; [2]甘肃农业大学动物医学院,甘肃兰州730070; [3]兰州大学基础医学院,甘肃兰州730030 |
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| 基金项目： | “十二五”国家科技支撑计划项目（2012BAD12B03）; 中央级科研单位基本科研业务费（1610322014028） |
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| 摘 要： | 目的：研发奶牛抗菌肽生物制剂,防治奶牛子宫内膜疾病。方法：采用酸性尿素聚丙烯酰胺凝胶电泳和琼脂糖弥散法检测子宫内膜组织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出相应的抗菌条带,HPLC进一步分离纯化,琼脂糖弥散法检测纯化物的抗菌活性,Tricine-SDS-PAGE电泳检测其分子量。结果：粗提物中有两种主要的抗菌成分,其对金黄色葡萄球菌26003、大肠杆菌BL-21、牛致病性大肠杆菌4605、绿脓杆菌10104具有抗菌活性。上层条带的主要色谱峰出现在43、45 min,其中45 min收集的样品有较强的抗菌活性,其相对分子质量约为14 k Da。下层条带主要色谱峰出现在5、35、43、45、46、47、50 min,其中35、43、45 min收集的样品有抗菌活性,其相对分子质量分别约为14-16、6和12 k Da。各个组分对大肠杆菌BL-21和金黄色葡萄球菌26003均具有抑菌活性。结论：从奶牛子宫内膜组织分离出了5个抗菌成分,从相对分子质量、抗菌活性等方面初步认定为抗菌多肽。
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| 关 键 词： | 子宫内膜 奶牛 抗菌肽 分离纯化 |
Isolation,purification and antimicrobial activity evaluation of antibacterial poiypeptide from uterus endometrial tissue of dairy cows |
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| Institution: | WANG Dong-sheng, MI Bao-ming, LI Xia, ZHANG Shi-dong, LI Shi-hong, ZONG Rui-qian, ZHU Bing-dong, WANG Gui-rong, YAN Zuo-ting (1, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS, Research Center of Clinical Veterinary Medicine of CAAS, Lanzhou Gansu 730050： 2.College of Veterinary Medicine, Gansu Agricultural University. Lanzhou Gansu 730070; 3. Basic Medical College of Lanzhou University, Lanzhou Gansu 730030, China) |
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| Abstract: | Objective： To research and develop biological agents of antimicrobial peptides for treatment of endometritis in dairy cows. Methods： The antibacterial activity of acid-soluble extracts from endometrial tissue was detected by acid urea PAGE and agarese diffusion assay. Anti-bacterial bands were isolated from gels with the application of agarhoresis and horizontal electrophoretic elution. Then, the separation and purification were further conducte antibacterial aetivity was detected by agarose diffusion assay and molecular weight was determined by Tr Results： There were two main components with anti-bacterial activity to S.aureus 26003, E. coil BL-21, bovicoli 4605 and Pseudomonas aeruginosa 10104. The main chromatographic peak of the upper bands appeared rain. The smnple at 45 rain present stronger anti-bacterial activity, and its molecular weight was about 1 chromatographic peak of the lower bands appeared at 5 rain, 35 rain, 43 rain, 45 rain, 46 min, 47 min and samples at 35 rain. 43 rain and 45 min showed anti-bacterial activity and molecular weight were about 14-16. 6 ,nd 12 kDa, respectively. These components had antibacterial activity on E, eoli and Staphylococcus aureus 26003 BL-2t. Conclusion： There were five components with antibacterial activities separated from dairy endometrial tissue. Although it also required further study, they were preliminarily considered as antimicrobial peptides according to the evaluation of the molecular weight and antihacterial activities. |
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| Keywords: | endometrial tissue： anti-microbial peptide separation and purification |
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