生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2013年
6期
753-758
,共6页
胡丹%张先云%耿美玲%张凤玉%郝丽娜%朱静%郑峰%潘秀珍%王长军
鬍丹%張先雲%耿美玲%張鳳玉%郝麗娜%硃靜%鄭峰%潘秀珍%王長軍
호단%장선운%경미령%장봉옥%학려나%주정%정봉%반수진%왕장군
2型猪链球菌%菌毛%SSU0473蛋白%原核表达%免疫保护
2型豬鏈毬菌%菌毛%SSU0473蛋白%原覈錶達%免疫保護
2형저련구균%균모%SSU0473단백%원핵표체%면역보호
Streptococcus suis serotype 2%pilus%protein SSU0473%prokaryotic expression%immunoprotection
目的:探究2型猪链球菌(S.suis2)强毒株05ZYH33的srtF基因簇编码的菌毛结构亚蛋白SSU0473的细菌定位及其免疫保护效能。方法:原核表达截短的SSU0473(tSSU0473),并以亲和层析法纯化目的蛋白,Western印迹检测tSSU0473蛋白的免疫原性,ELISA法检测多抗血清的效价及IgG亚型,小鼠试验测试重组蛋白的免疫保护效能,免疫电镜观测tSSU0473蛋白的细菌定位。结果:在原核系统中表达了tSSU0473蛋白;ELISA结果显示重组蛋白能够刺激小鼠产生高效价的免疫抗体;动物试验表明tSSU0473蛋白免疫小鼠可抵御致死剂量病原体的攻击,显示出较好的免疫保护作用;免疫电镜检测显示tSSU0473蛋白定位于细菌表面。结论:菌毛亚蛋白tSSU0473是S.suis2膜表面蛋白,具有良好的免疫原性和免疫保护性,可作为S.suis2亚单位疫苗的候选分子。研究结果为系统揭示S.suis2的菌毛生物学结构与功能奠定了基础。
目的:探究2型豬鏈毬菌(S.suis2)彊毒株05ZYH33的srtF基因簇編碼的菌毛結構亞蛋白SSU0473的細菌定位及其免疫保護效能。方法:原覈錶達截短的SSU0473(tSSU0473),併以親和層析法純化目的蛋白,Western印跡檢測tSSU0473蛋白的免疫原性,ELISA法檢測多抗血清的效價及IgG亞型,小鼠試驗測試重組蛋白的免疫保護效能,免疫電鏡觀測tSSU0473蛋白的細菌定位。結果:在原覈繫統中錶達瞭tSSU0473蛋白;ELISA結果顯示重組蛋白能夠刺激小鼠產生高效價的免疫抗體;動物試驗錶明tSSU0473蛋白免疫小鼠可牴禦緻死劑量病原體的攻擊,顯示齣較好的免疫保護作用;免疫電鏡檢測顯示tSSU0473蛋白定位于細菌錶麵。結論:菌毛亞蛋白tSSU0473是S.suis2膜錶麵蛋白,具有良好的免疫原性和免疫保護性,可作為S.suis2亞單位疫苗的候選分子。研究結果為繫統揭示S.suis2的菌毛生物學結構與功能奠定瞭基礎。
목적:탐구2형저련구균(S.suis2)강독주05ZYH33적srtF기인족편마적균모결구아단백SSU0473적세균정위급기면역보호효능。방법:원핵표체절단적SSU0473(tSSU0473),병이친화층석법순화목적단백,Western인적검측tSSU0473단백적면역원성,ELISA법검측다항혈청적효개급IgG아형,소서시험측시중조단백적면역보호효능,면역전경관측tSSU0473단백적세균정위。결과:재원핵계통중표체료tSSU0473단백;ELISA결과현시중조단백능구자격소서산생고효개적면역항체;동물시험표명tSSU0473단백면역소서가저어치사제량병원체적공격,현시출교호적면역보호작용;면역전경검측현시tSSU0473단백정위우세균표면。결론:균모아단백tSSU0473시S.suis2막표면단백,구유량호적면역원성화면역보호성,가작위S.suis2아단위역묘적후선분자。연구결과위계통게시S.suis2적균모생물학결구여공능전정료기출。
Objective: To elucidate the inmmunoprotection of pilus subunit SSU0473 encoded by the srtF pilus is-land in the Chinese strain 05ZYH33 of Streptococcus suis serotype 2(S.suis2). Methods: The truncated protein tSSU0473 was expressed in prokaryotic cells and was purified by affinity chromatography. Then recombinant pro-tein was analyzed by Western blotting, and the anti-SSU0473 serum was prepared by immunizing mice with recom-binant tSSU0473 protein and analyzed by ELISA. The location site of tSSU0447 protein in bacterium was detected by immune electron microscopy. Results: The tSSU0473 protein was expressed successfully in prokaryotic system. The high titer of the serum was obtained, ELISA result indicated that the recombinant protein can react with mice anti-SSU0473 serum. Animal test showed that vaccinating mice with recombinant tSSU0473 protein conferred a significant protection. The tSSU0473 protein was detected on bacterial surface by immune electron microscopy. Conclusion: Protein tSSU0473 could serve as a vaccine candidate of S.suis2, and provides the foundation for sys-tematically illustrating the role that pilus island plays in pathogenicity of S.suis2.