影响坏死梭杆菌分泌白细胞毒素因素的研究  

Study on Factors Influencing the Leukotoxin Secretion by F. necrophorum

作　　者：冯二凯[1] 陈立志[1] 刘晓颖[1] 汪孙杰[1,2] 曹阅[1] 徐晶[1,2] 

机构地区：[1]中国农业科学院特产研究所特种经济动物分子生物学省部共建国家重点实验室,吉林吉林132109 [2]江苏科技大学生物与环境工程学院,江苏镇江212018

出　　处：《动物医学进展》2012年第11期12-17,共6页Progress In Veterinary Medicine

基　　金：中央级公益性科研院所基本科研院所专项资金(2012ZL087);吉林省科技发展计划项目(201205028);吉林市科技支撑计划项目(201262509)

摘　　要：利用已分离坏死梭杆菌菌株,建立坏死梭杆菌白细胞毒素活性体外检测方法,并评价培养条件对坏死梭杆菌天然白细胞毒素活性的影响。结果表明,预还原、厌氧无菌心脑浸液肉汤培养基(pH7.3左右)、培养时间10h～12h(对数生长期后期),此工艺参数下获取的细菌培养物上清液的白细胞毒性最大。获得的坏死梭杆菌天然白细胞毒素经Western blot证实,能被抗白细胞毒素重组BSBSE蛋白的兔血清识别,表明提取的白细胞毒素具有反应原性。In order to evaluate the effect of culture conditions on leukotoxin secretion by Fusobacterium necrophorum（F.necrophorum）,a cytotoxic test for assessing the leukotoxicity of F.necrophorum culture supernatant was established,which employed the polymorph nuclear leukocytes（PMNs） as the target cells.And then,the influence of pH,media,culture time and biotype of bacterium on leukotoxin secretion by F.necrophorum was further studied.The results indicated that the maximal leukotoxin activity was observed when F.necrophorum（biotype A） was grown in pre-reduced anaerobically sterilized brain heart infusion（PRAS-BHI） broth media at a pH of 7.2-7.5 for about 10-12hours.At the end of culturing,ultrafiltration was used to separate the leukotoxin from the culture supernatant of F.necrophorum,whereupon the vaccine is produced by inactivation of crude leukotoxin.Western blot indicated that the crude leukotoxin could be recognized by the rabbit serum that raised against the recombinant proteins of BSBSE of FN（4） strain of F.necrophorum.

关 键 词：坏死梭杆菌 白细胞毒素 分泌 影响 

分 类 号：S852.616.3[农业科学—基础兽医学]