问号澳洲型钩端螺旋体复合抗原基因ompL_1/flaB_2表达质粒的构建

Construction of expression vector including the multi-gene encoding ompL_1 and flaB_2 from Leptospira interrogans serovar australis

目的 为增强澳洲型钩端螺旋体 (简称澳洲型钩体 ) 6 0 7株抗原的免疫原性 ,而构建其外膜蛋白抗原基因ompL1和内鞭毛抗原基因flaB2 的复合抗原基因重组质粒。方法 通过聚合酶链反应扩增ompL1及flaB2 ,将其分别克隆到pcD NA3 1/Myc-His(+)载体T7启动子下游 ,进行序列测定分析 ,在此基础上将ompL1及flaB2 同时克隆至表达载体 pcD NA3 1/Myc -His(+) ,构建成A - pcLMF1复合抗原基因表达质粒。结果 澳洲型钩体 6 0 7株的ompL1及flaB2 与报道的其它株钩体的序列呈很高的同源性。澳洲型钩体重组质粒A -pcLMF1经酶切鉴定证实 :有一 1 8kb片段插入。结论 澳洲型钩体复合抗原基因ompL1/flaB2

Aim To construct a multi antigen gene recombinant plasmid including an outer membrane protein gene(ompL 1) and a flagellin B gene(flaB 2) to enhance the protective immunity of leptospira interrogans serovar australis strain 607 Methods The DNA fragments encoding ompL 1 and flaB 2 were amplified respectively by PCR,then they were cloned separately into the down stream of the T7 promoter of vector pcDNA3 1/Myc-His(+),and analyzed by DNA sequencing On the basis,the both gene were cloned into the same vector pcDNA 3 1/Myc-His(+),A-pcLMF1 recombinant plasmid of multi antigen gene was constructed Result Analysis of the DNA sequence of ompL 1 and flaB 2 from L australis strain 607 showed that the sequences were high level identity with other strains Conclusions A fusion expression plasmid containing multi antigenic gene ompL 1/flaB 2 from L australis was constructed