PurC影響馬鏈球菌獸疫亞種致腦膜炎的作用機(jī)制
發(fā)布時(shí)間:2021-12-29 04:15
馬鏈球菌獸疫亞種(Streptococcus equi ssp.zooepidemicus,SEZ)屬蘭氏分群的C群β溶血鏈球菌,可引起敗血癥,關(guān)節(jié)炎,心內(nèi)膜炎和腦膜炎。其宿主譜較廣,可感染豬、馬、犬、貓等多種與人類關(guān)系密切的動(dòng)物。在我國(guó),SEZ是造成豬鏈球菌病的重要豬源病原體,是引起豬細(xì)菌性腦膜炎的主要病原之一。同時(shí),全球范圍內(nèi)已有大量人類感染SEZ引起腦膜炎的病例報(bào)道,大多數(shù)感染與密切接觸患病動(dòng)物或食用被污染的食物有關(guān),嚴(yán)重時(shí)可導(dǎo)致死亡。因此,SEZ人畜共感染和致腦膜炎的特性已對(duì)人類健康構(gòu)成巨大威脅。本文對(duì)PurC影響馬鏈球菌獸疫亞種ATCC35246株致腦膜炎相關(guān)機(jī)制進(jìn)行了研究。并且篩選了其他可能破壞血腦屏障的SEZ分泌蛋白。以上研究結(jié)果為進(jìn)一步闡明SEZ引起腦膜炎的機(jī)制提供理論依據(jù)。1、馬鏈球菌獸疫亞種PurC與HEK 293T細(xì)胞互作蛋白的篩選PurC(磷;被溥-琥珀酰基酰胺合成酶,SAICAR合成酶)為參與嘌呤生物合成途徑第七步的催化酶。課題組前期對(duì)SEZ ATCC35246強(qiáng)毒株的全基因組進(jìn)行比較基因組學(xué)分析,在其II號(hào)毒力島中發(fā)現(xiàn)了 SEZ ATCC35246特有...
【文章來源】:南京農(nóng)業(yè)大學(xué)江蘇省 211工程院校 教育部直屬院校
【文章頁數(shù)】:172 頁
【學(xué)位級(jí)別】:博士
【部分圖文】:
圖2-1?PCR及雙酶切鑒定purC-pAcGFP載體構(gòu)建??Fiure?2-1.Detection?ofurC-AcGFPlasmid??
?第二章馬鏈球菌獸疫亞種PurC與HEK293T細(xì)胞互作蛋白的篩選???A?PuiC-OFP?pAeGFP?B?M?12?3???—?PU[C.(肝??■■??圖2-2?Pu「C細(xì)胞內(nèi)轉(zhuǎn)染及表達(dá)檢測(cè)??Figure?2-2?PurC?transfection?and?intracellular?expression?detection??(A)?Observation?of?intracellular?expression?by?fluorescence?microscopy;?(B)?Western?blot?was?used?to?detect?the?expression?of?plasmid?in??cells.?Templet?in?lane?1-2:?Cell?lysate?transfected?with?pwrC-pAcGFP?plasmid;Templet?in?lane?3:?Cell?lysate?transfected?with?pAeGFP?vector;??M:?protein?marker,?the?red?band?represents?70?kDa.??2.3?SILAC方法篩選HEK239T細(xì)胞中與PurC互作的蛋白??用;wC-pAcGFP和pAeGFP質(zhì)粒分別轉(zhuǎn)染Heavy和Light標(biāo)記的HEK293T細(xì)胞??中。轉(zhuǎn)染后的細(xì)胞總蛋白1:1混合,用小鼠anti-GFP單克隆抗體CO-IP富集,獲得的??蛋白用胰酶消化并通過LC-MS/MS分析(如圖2-3所示)。??pAcGFP-parC?pAeGFP??〇?o??Hwvy?l*b?。浚?2ft3T?c*M?Tr?n?
?第二章馬鏈球菌獸疫亞種PurC與HEK293T細(xì)胞互作蛋白的篩選???A?PuiC-OFP?pAeGFP?B?M?12?3???—?PU[C.(肝??■■??圖2-2?Pu「C細(xì)胞內(nèi)轉(zhuǎn)染及表達(dá)檢測(cè)??Figure?2-2?PurC?transfection?and?intracellular?expression?detection??(A)?Observation?of?intracellular?expression?by?fluorescence?microscopy;?(B)?Western?blot?was?used?to?detect?the?expression?of?plasmid?in??cells.?Templet?in?lane?1-2:?Cell?lysate?transfected?with?pwrC-pAcGFP?plasmid;Templet?in?lane?3:?Cell?lysate?transfected?with?pAeGFP?vector;??M:?protein?marker,?the?red?band?represents?70?kDa.??2.3?SILAC方法篩選HEK239T細(xì)胞中與PurC互作的蛋白??用;wC-pAcGFP和pAeGFP質(zhì)粒分別轉(zhuǎn)染Heavy和Light標(biāo)記的HEK293T細(xì)胞??中。轉(zhuǎn)染后的細(xì)胞總蛋白1:1混合,用小鼠anti-GFP單克隆抗體CO-IP富集,獲得的??蛋白用胰酶消化并通過LC-MS/MS分析(如圖2-3所示)。??pAcGFP-parC?pAeGFP??〇?o??Hwvy?l*b?。浚?2ft3T?c*M?Tr?n?
【參考文獻(xiàn)】:
期刊論文
[1]豬鏈球菌病與豬鏈球菌2型[J]. 陸承平. 科技導(dǎo)報(bào). 2005(09)
本文編號(hào):3555383
【文章來源】:南京農(nóng)業(yè)大學(xué)江蘇省 211工程院校 教育部直屬院校
【文章頁數(shù)】:172 頁
【學(xué)位級(jí)別】:博士
【部分圖文】:
圖2-1?PCR及雙酶切鑒定purC-pAcGFP載體構(gòu)建??Fiure?2-1.Detection?ofurC-AcGFPlasmid??
?第二章馬鏈球菌獸疫亞種PurC與HEK293T細(xì)胞互作蛋白的篩選???A?PuiC-OFP?pAeGFP?B?M?12?3???—?PU[C.(肝??■■??圖2-2?Pu「C細(xì)胞內(nèi)轉(zhuǎn)染及表達(dá)檢測(cè)??Figure?2-2?PurC?transfection?and?intracellular?expression?detection??(A)?Observation?of?intracellular?expression?by?fluorescence?microscopy;?(B)?Western?blot?was?used?to?detect?the?expression?of?plasmid?in??cells.?Templet?in?lane?1-2:?Cell?lysate?transfected?with?pwrC-pAcGFP?plasmid;Templet?in?lane?3:?Cell?lysate?transfected?with?pAeGFP?vector;??M:?protein?marker,?the?red?band?represents?70?kDa.??2.3?SILAC方法篩選HEK239T細(xì)胞中與PurC互作的蛋白??用;wC-pAcGFP和pAeGFP質(zhì)粒分別轉(zhuǎn)染Heavy和Light標(biāo)記的HEK293T細(xì)胞??中。轉(zhuǎn)染后的細(xì)胞總蛋白1:1混合,用小鼠anti-GFP單克隆抗體CO-IP富集,獲得的??蛋白用胰酶消化并通過LC-MS/MS分析(如圖2-3所示)。??pAcGFP-parC?pAeGFP??〇?o??Hwvy?l*b?。浚?2ft3T?c*M?Tr?n?
?第二章馬鏈球菌獸疫亞種PurC與HEK293T細(xì)胞互作蛋白的篩選???A?PuiC-OFP?pAeGFP?B?M?12?3???—?PU[C.(肝??■■??圖2-2?Pu「C細(xì)胞內(nèi)轉(zhuǎn)染及表達(dá)檢測(cè)??Figure?2-2?PurC?transfection?and?intracellular?expression?detection??(A)?Observation?of?intracellular?expression?by?fluorescence?microscopy;?(B)?Western?blot?was?used?to?detect?the?expression?of?plasmid?in??cells.?Templet?in?lane?1-2:?Cell?lysate?transfected?with?pwrC-pAcGFP?plasmid;Templet?in?lane?3:?Cell?lysate?transfected?with?pAeGFP?vector;??M:?protein?marker,?the?red?band?represents?70?kDa.??2.3?SILAC方法篩選HEK239T細(xì)胞中與PurC互作的蛋白??用;wC-pAcGFP和pAeGFP質(zhì)粒分別轉(zhuǎn)染Heavy和Light標(biāo)記的HEK293T細(xì)胞??中。轉(zhuǎn)染后的細(xì)胞總蛋白1:1混合,用小鼠anti-GFP單克隆抗體CO-IP富集,獲得的??蛋白用胰酶消化并通過LC-MS/MS分析(如圖2-3所示)。??pAcGFP-parC?pAeGFP??〇?o??Hwvy?l*b?。浚?2ft3T?c*M?Tr?n?
【參考文獻(xiàn)】:
期刊論文
[1]豬鏈球菌病與豬鏈球菌2型[J]. 陸承平. 科技導(dǎo)報(bào). 2005(09)
本文編號(hào):3555383
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