發(fā)布時(shí)間：2019-07-02 14:35

【摘要】：為了研究豬鏈球菌2型假定外膜蛋白Omp40的免疫保護(hù)性作用,根據(jù)已發(fā)表的豬鏈球菌2型豬源98HAH33株的Omp40蛋白的基因序列,選擇免疫原性較好的基因序列設(shè)計(jì)并合成引物,以HA9801株的基因組DNA為模板,采用PCR擴(kuò)增出Omp40蛋白的部分基因定向克隆至表達(dá)載體pET-32a(+)中并轉(zhuǎn)化入BL21(DE3)宿主菌。重組菌經(jīng)IPTG誘導(dǎo)后的SDS-PAGE圖譜顯示在52.5 ku處出現(xiàn)融合蛋白的條帶。Western-blot結(jié)果顯示,其與豬源ZY05719株康復(fù)血清有良好的免疫反應(yīng)性。動(dòng)物試驗(yàn)結(jié)果證實(shí),經(jīng)Omp40蛋白免疫小鼠的血清抗體效價(jià)在二免后第7天高達(dá)1∶50 800,IL-4和IFN-γ mRNA的表達(dá)量在二免后第7天均與對(duì)照組相比差異顯著(P0.05),并對(duì)豬鏈球菌2型強(qiáng)致病性株HA9801感染斑馬魚提供22%的免疫保護(hù)率。本研究為今后豬鏈球菌病亞單位疫苗的研究提供了實(shí)驗(yàn)依據(jù)。
[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，

本文編號(hào)：2509010