Position: Home > Articles > Expression and antigenicity analysis of hyaluronidase (HylB) of piscine Streptococcus agalactiae
Animal Husbandry & Veterinary Medicine
2014,46
(1)
24-27
鱼源无乳链球菌透明质酸酶的表达及抗原性分析
作 者：
王兆飞;郭长明;陆承平;刘永杰
单 位：
南京农业大学动物医学院
关键词：
鱼源无乳链球菌;透明质酸酶;hylB;表达;免疫原性
摘 要：
透明质酸酶(HylB)是无乳链球菌重要的毒力因子。根据本实验室分离并公布在GenBank上的罗非鱼源无乳链球菌GD201008-001全基因组序列,PCR扩增获得长为2 346 bp,覆盖hylB保守功能区的基因片段。将该片段采用限制性内切酶BamHⅠ和XhoⅠ消化后亚克隆至表达载体pET28a(+),经双酶切、测序鉴定后转化至大肠杆菌BL21(DE3)中,进行体外原核表达,获得88.5 ku的目的蛋白。将纯化后的目的蛋白免疫家兔制备多克隆抗体,经ELISA和Western blot鉴定该蛋白具有良好的免疫原性。
译 名：
Expression and antigenicity analysis of hyaluronidase (HylB) of piscine Streptococcus agalactiae
作 者：
WANG Zhaofei;GUO Changming;LU Chengping;LIU Yongjie;College of Veterinary Medicine,Nanjing Agricultural University;
关键词：
piscine Streptococcus agalactiae;;hyaluronidase;;hylB;;expression;;antigenicity
摘 要：
Hyaluronidase( HylB) is an important virulence factor of Streptococcus agalactiae. The hylB gene was amplified from the ge- nomic DNA of S. agalactiae strain GD201008- 001 isolated from Tilapia. A 2 346 bp gene fragment encoding the conserved functional pro- tein domain was subcloned into pET28a( +) vector digested by BamHⅠand XhoⅠ enzymes. And then the recombinant plasmid was trans- formed into BL21( DE3) and induced to express by IPTG. SDS- PAGE analysis showed that a specific band of protein with the molecular weight of 88. 5 ku was obtained. Further the recombinant protein was intracutaneously inoculated into rabbits and the high titre of antiserum was obtained. Western blot analysis revealed that the rabbit antiserum against HylB could react with the whole proteins from S. agalactiae. It suggests that the recombinant protein has good antigenicity.
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