Molecular Cloning, expression and purification of immunogenic proteins Pgi and Spb1 of Streptococcus agalactiae
作者单位：Key Laboratory for Aquatic Products Safety of Ministry of Education School of Life Sciences Sun Yat-sen University
会议名称：《中国水产学会鱼病专业委员会2013年学术研讨会》
会议日期：2013年
关 键 词：Streptococcus agalactiae Pgi Spb1 vaccine
摘 要：Streptococcus agalactiae,also known as Group B Streptococcus(GBS),is a majorpathogen that causes serious infections in numerous species of freshwater and marine *** antigens have potentials to be ideal vaccines against GBS *** thisstudy,we selected 2 immunogenic proteins glucose-6-phosphate isomeras(Pgi)and PI-2bBackbone protein(Spb1)to test their potentials as *** we extracted *** DNA to amplify the ORF of pgi and spb1 by PCR,and clone to pET-28a(+)expressionvector by T4 ligase to construct recombination expression *** plasmids weretransformed to *** BL21(DH3)and induced to express by *** showed the MW ofrPgi was0 kDa,and rSpb1 was3 *** 16℃,0.2mM IPTG,both of them existed in thesupernatant and *** the following vaccine preparation,we chose the soluble partsto conduct purification on Ni-NTA *** proteins were desperately eluted with 250 mM and300mM imidazole,and ultra-filtered to remove *** concentration of rPgi and rSpb1were diluted to 1mg/ml in PBS and mixed as 1:1 emulsions in adjuvant to formulate *** task is to test the protection rate offered by the proteins in Nile tilapia against ***.