豚链球菌和停乳链球菌类M蛋白及其抗原性分析  

M-like proteins and antigenicity of M-like proteins in Streptococcus iniae and Streptococcus dysgalactiae

作　　者：汪笑宇[1] 战文斌[1] 邢婧[1] 绳秀珍[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003

出　　处：《水产学报》2008年第6期945-949,共5页Journal of Fisheries of China

基　　金：国家自然科学基金(30771648);国家"八六三"高技术研究发展计划基金(2006AA100306)

摘　　要：分别用热酸抽提法、碱抽提法和胃蛋白酶消化法分离豚链球菌NUF849的类M蛋白,通过SDS-PAGE和Western-blotting印迹法分析比较三种方法所提类M蛋白的组成和抗原性差异。结果表明,热酸抽提法分离制备的类M蛋白得率和纯度都较其它两种方法高,蛋白的抗原性保持较好。采用热酸抽提法提取5株豚链球菌NUF633、NUF693、NUF701、NUF812、NUF849和2株停乳链球菌SD和L2的类M蛋白,其SDS-PAGE图谱明显不同,种间差异明显,豚链球菌主要类M蛋白的分子量分别为60、48、37、30、27、24和15ku,停乳链球菌主要类M蛋白的分子量分别为68、48、42、37、29、26、23、21、19、17、15、14和11ku。Western-blotting结果显示,兔抗豚链球菌血清可与豚链球菌的两条类M蛋白带结合,分子量分别为60和37ku;与停乳链球菌的两条类M蛋白带结合,分子量为68和37ku。两种链球菌主要类M蛋白组成及其抗原性差异明显。M-like protein was reported mostly in Streptococcus equi and S. suis, and was a kind of important antigen of Streptococcus. In order to analyze antigenicity of M-like protein in S. iniae, three different extraction methods （hot acid extraction, alkaline extraction and pepsin digestion） were used for preparing M-like proteins from Streptococcus iniae and Streptococcus dysgalactiae. The M-like proteins of the two streptococci were analyzed by using SDS-PAGE and Western-blotting. By comparing the results of the three methods, the M-like proteins extracted by hot acid had the strongest immunogenicity. The SDS-PAGE diagrams of the M-like proteins of the two streptococci were distinctly different. The M-like proteins extracted from S. iniae contained 7 minor protein bands with molecular weights of 60, 48, 37, 33, 27, 24 and 15 ku, the M-like proteins extracted from S. dysgalactiae contained 2 minor protein bands with molecular weights of 68 and 42 ku. After SDS- PAGE, in the Western-blotting with rabbit antiserum against S. iniae whole cells, only 2 M-like proteins of S. iniae with molecular weights of 60 ku and 37 ku were detected, and only 2 M-like proteins of S. dysgalactiae with molecular weights of 68 ku and 37 ku. It showed that the M-like proteins of the two streptococci had different composition and antigenicity.

关 键 词：豚链球菌 停乳链球菌 类M蛋白 抗原性 

分 类 号：S948[农业科学—水产养殖]