红斑丹毒丝菌免疫原性蛋白的鉴定及其编码基因的克隆和测序

发布时间：2018-03-03 22:14

本文选题：红斑丹毒丝菌　切入点：NaOH提取抗原　出处：《微生物学报》2016年02期 　论文类型：期刊论文

【摘要】：【目的】为深入研究红斑丹毒丝菌的免疫保护性抗原及其致病机制,采用免疫蛋白组学技术鉴定红斑丹毒丝菌的免疫原性蛋白。【方法】通过SDS-PAGE电泳分离红斑丹毒丝菌C43065株的NaOH提取抗原,用兔抗NaOH提取抗原抗血清经Western blot检测免疫原性蛋白,通过MALDI-TOF质谱技术鉴定蛋白种类,并对部分免疫原性蛋白的编码基因进行克隆和测序。【结果】通过MALDI-TOF质谱技术从C43065株NaOH提取抗原中鉴定出9个免疫原性蛋白,分别为Spa A、伴侣蛋白GroEL、烯醇化酶、ATP结合盒转运蛋白、丙酮酸脱氢酶复合物E1、甘油醛-3-磷酸脱氢酶、果糖二磷酸醛缩酶、50S核糖体蛋白L1、30S核糖体蛋白S4。其中烯醇化酶、ATP结合盒转运蛋白、甘油醛-3-磷酸脱氢酶和果糖二磷酸醛缩酶已被证实与链球菌、牙龈卟啉单胞菌、脑膜炎奈瑟菌和结核分枝杆菌的致病性相关。C43065株伴侣蛋白GroEL、烯醇化酶、ATP结合盒转运蛋白、丙酮酸脱氢酶复合物E1、甘油醛-3-磷酸脱氢酶和果糖二磷酸醛缩酶编码基因大小分别为1614、1296、1260、1005和867 bp,与已公布的红斑丹毒丝菌Fujisawa株相应基因的相似度高达98%。【结论】本文所鉴定的9个免疫原性蛋白,为进一步开展红斑丹毒丝菌保护性抗原及其致病机制研究奠定基础。
[Abstract]:[objective] to study the immune protective antigen of erysipelas erythematosus and its pathogenic mechanism. The immunogenicity protein of erysipelas erythematosus was identified by immunoproteomics technique. [methods] the NaOH antigen of Rhizoma erythematosus C43065 strain was isolated by SDS-PAGE electrophoresis, and the immunogenicity protein was detected by Western blot in rabbit anti-#en2# antiserum. The protein types were identified by MALDI-TOF mass spectrometry, and the coding genes of some immunogenic proteins were cloned and sequenced. [results] 9 immunogenic proteins were identified from C43065 strain NaOH by MALDI-TOF mass spectrometry. They are Spa A, chaperone protein Groel, enolase ATPase ATP-binding cassette transporter, pyruvate dehydrogenase complex E1, glyceraldehyde 3-phosphate dehydrogenase. Fructose diphosphate aldolase 50s ribosomal protein L1 / 30s ribosomal protein S4. among them enolase ATP binding cassette transporter, glyceraldehyde 3-phosphate dehydrogenase and fructose diphosphate aldolase have been confirmed to be associated with Streptococcus gingivalis, porphyromonas gingivalis. Pathogenicity of Neisseria meningitidis and Mycobacterium tuberculosis. The coding genes of pyruvate dehydrogenase complex E1, glyceraldehyde 3-phosphate dehydrogenase and fructose diphosphate aldolase were 1614, 1296, 1260,1005 and 867 BP, respectively. The similarity of the genes with the published Fujisawa strain was 98. [conclusion]. Nine immunogenic proteins were identified. It lays a foundation for further research on protective antigen and pathogenic mechanism of erysipelas erythematosus.
【作者单位】：伊犁师范学院生物与地理科学学院;吉首大学生物资源与环境科学学院;中国疾病预防控制中心传染病预防控制所;
【基金】：国家自然科学基金项目(31360613,31072142)~~
【分类号】：S852.6

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