猪链球菌2型假定外膜蛋白Omp40免疫原性的鉴定
发布时间：2019-07-02 14:35
【摘要】：为了研究猪链球菌2型假定外膜蛋白Omp40的免疫保护性作用,根据已发表的猪链球菌2型猪源98HAH33株的Omp40蛋白的基因序列,选择免疫原性较好的基因序列设计并合成引物,以HA9801株的基因组DNA为模板,采用PCR扩增出Omp40蛋白的部分基因定向克隆至表达载体pET-32a(+)中并转化入BL21(DE3)宿主菌。重组菌经IPTG诱导后的SDS-PAGE图谱显示在52.5 ku处出现融合蛋白的条带。Western-blot结果显示,其与猪源ZY05719株康复血清有良好的免疫反应性。动物试验结果证实,经Omp40蛋白免疫小鼠的血清抗体效价在二免后第7天高达1∶50 800,IL-4和IFN-γ mRNA的表达量在二免后第7天均与对照组相比差异显著(P0.05),并对猪链球菌2型强致病性株HA9801感染斑马鱼提供22%的免疫保护率。本研究为今后猪链球菌病亚单位疫苗的研究提供了实验依据。
[Abstract]:In order to study the immune protective effect of Streptococcus suis type 2 assumed outer membrane protein Omp40, according to the published gene sequence of Omp40 protein of Streptococcus suis type 2 98HAH33 strain, primers were designed and synthesized by selecting the gene sequence with good immunogenicity. Using the genomic DNA of HA9801 strain as template, some genes of Omp40 protein amplified by PCR were cloned into expression vector pET-32a () and transformed into BL21 (DE3) host strain. The SDS-PAGE map of the recombinant strain induced by IPTG showed that the fusion protein band appeared at 52.5 ku. Western-blot results showed that the recombinant strain had good immunoreactivity with the rehabilitation serum of pig ZY05719 strain. The results of animal test showed that the serum antibody titer of mice immunized with Omp40 protein was as high as 1:50 800 on the 7th day after second immunization, and the expression of IL 鈮，
本文编号：2509010
[Abstract]:In order to study the immune protective effect of Streptococcus suis type 2 assumed outer membrane protein Omp40, according to the published gene sequence of Omp40 protein of Streptococcus suis type 2 98HAH33 strain, primers were designed and synthesized by selecting the gene sequence with good immunogenicity. Using the genomic DNA of HA9801 strain as template, some genes of Omp40 protein amplified by PCR were cloned into expression vector pET-32a () and transformed into BL21 (DE3) host strain. The SDS-PAGE map of the recombinant strain induced by IPTG showed that the fusion protein band appeared at 52.5 ku. Western-blot results showed that the recombinant strain had good immunoreactivity with the rehabilitation serum of pig ZY05719 strain. The results of animal test showed that the serum antibody titer of mice immunized with Omp40 protein was as high as 1:50 800 on the 7th day after second immunization, and the expression of IL 鈮，
本文编号：2509010
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