畜牧与兽医2024，Vol.56Issue(1)：39-45,7.
大片吸虫磷酸丙糖异构酶基因的克隆表达和免疫原性分析
Cloning,expression and immunogenicity of triosephosphate isomerase of Fasciola gigantica
摘要
Abstract
Triosephosphate isomerase(TPI)is a key glycolytic enzyme that exists in the organisms and plays an important role in obtaining energy for the life of Fasciola gigantica.In this study,the full-length cDNA encoding FgTPI was cloned according to the gene sequence of TPI from Fasciola gigantica,and the primer with Bam HⅠ and XhoⅠ as enzyme digestion sites was designed.The cDNA from Fasciola gigantica was used as a template.The results were as follows:The open reading frame(ORF)of FgTPI was 762 bp,encoding 253 amino acids with an isoelectric point of 8.07.The ORF of was sub-cloned into pET-28a(+)vectors and expressed in Escherichia coli with a mo-lecular weight of 32 kDa.The BALB/c mouse was injected with the purified recombinant of FgTPI,and the polyclonal antibodies were pre-pared.ELISA assay analysis showed that rFgTPI could induce high levels of anti-rFgTPI-specific IgG antibodies.The result of western blot-ting showed that recombinant FgTPI had good antigenicity by using the serum from the sheep infected with Fasciola gigantica.The recombi-nant protein showed the optimal activity of TPI at pH 7.5 and temperature of 35℃,according to α-glycerol phosphate dehydrogenase cou-pling method.The above results indicated that FgTPI could be a potential candidate for vaccine against Fascioliasis,which would provide ba-sis for further investigation into the biological function of FgTPI.关键词
大片吸虫/磷酸丙糖异构酶/克隆/表达/酶活性分析Key words
Fasciola gigantica/triosephosphate isomerase/cloning/expression/enzymatic activity analysis分类
农业科技引用本文复制引用
姚文浩,吴兴隆,秦志博,李文豪,朱惠丽,王磊,胡建和,韩艳辉..大片吸虫磷酸丙糖异构酶基因的克隆表达和免疫原性分析[J].畜牧与兽医,2024,56(1):39-45,7.基金项目
国家重点研发计划项目(2021YFD1301200) （2021YFD1301200）
中原科技创新领军人才项目(224200510024) （224200510024）
