SmB/B’抗原表位的表达及其免疫原性研究

Study on the Expression and Immunogenicity of SmB/B' Epitope

为了构建SmB/B’抗原表位的真核表达载体 ,通过体内、外进行表达并研究其免疫原性 ,采用异硫氰酸胍一步法提取BALB/c小鼠脾脏总RNA ,通过RT PCR方法克隆了含编码SmB/B’抗原表位的目的DNA ,构建了其真核表达质粒pcDNA3 fSmB/B’ ,体外转染HeLa细胞检测其表达及与抗体结合活性。体内采用基因免疫的方法对其免疫原性进行研究。结果 :成功构建pcDNA3 fSmB/B’真核表达质粒 ,Westernblot显示其在真核细胞内可有效表达且与抗SmB/B’抗体具有结合活性 ,经基因免疫实验组小鼠皆产生抗SmB/B’抗体。构建的SmB/B’抗原表位真核表达载体可在体内、外有效表达 。

To construct the eukaryotic expression vector of SmB/B' epitope and to study its immunogenicity after its expression in vivo and in vitro,the target DNA encoding the SmB/B' epitope was cloned by RT PCR,and then the eukaryotic expression vector pcDNA3 f SmB/B' was constructed Its expression and antibody binding activities were detected in vitro,and its immunogenicity was determined by gene immunization in vivo It was found that the eukaryotic expression vector pcDNA3 fSmB/B' was constructed correctly,and it could be expressed in vitro with binding activity with the anti SmB/B' antibody All BALB/c mice immunized with this plasmid DNA produced anti SmB/B' antibody The experimental results indicate that the constructed eukaryotic expression vector of SmB/B' epitope can efficiently express in vitro and in vivo and the expressed epitope has immunogenicity