摘要
目的 克隆出血性大肠杆菌O157:H7的Stx2毒素B亚单位基因 ,进行序列分析并构建表达载体。方法 利用PCR技术从出血性大肠杆菌O157:H7染色体基因组中扩增出Stx2毒素B亚单位基因 ,并连接至质粒载体上 ,进行序列测定和分析 ,亚克隆构建表达载体 ,经SDS PAGE电泳分析目的蛋白表达量。结果 所克隆的基因序列与Genbank上基因序列高度同源 ,所表达目的蛋白约占宿主菌蛋白的 2 0 %。
Objective To clone,sequence and construct an expression vector of Stx 2 toxin subunit B gene of EHEC O 157 :H 7 .Methods Amplify Stx 2 toxin subunit B gene from the genomic DNA of EHEC O 157 :H 7 strain by PCR,clone into plasmid pUC19,sequence and digest with restriction endonuclease,then subclone into plasmid pET28a and transform to BL21(DE3)for expression.Determine the expression level of goal protein by SDS PAGE.Results The sequence of cloned gene was highly homologous to that in Genbank,and the expressed product contained about 20% of host sometic protein.Conclusion The study laid a foundation of further development of vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2004年第1期16-17,22,共3页
Chinese Journal of Biologicals
