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中文PI字克雷伯氏肺炎U菌v膜多醣噬菌w
英文PI字Klebsiellapneumoniaecapsularpolysaccharidesbacteriophage
W科e分W科e＞tWc生命科W＞微生物W
中文摘要^去二十年恚谔酬m出F新型侵u性克雷伯氏肺炎U菌病例。此菌斐稍l性肝、⊙孕菘恕⒕⑥D移性眼妊住⒛X膜炎等乐夭♂纭？死撞戏窝U菌菌w最外拥那v膜c其黏附、毒性有O大P，特e是毒性^的K1和K2v膜血清型菌株。施打疫苗是疾病A防重要策略之一。市售肺炎球菌疫苗即檠}捣Nv膜多醣寡分子（capsularpolysaccharideoligomers）混合成多效r疫苗。故ふ夷蚯懈罹薮笄v膜成小分子，又不影其免疫原特性之方法是我努力的目恕Ｒ延醒芯恐赋鍪删w能ηv膜多醣a生切割降解效果（CPSdepolymerization）。因此，在本研究中，我收取未淼脑c克雷伯氏肺炎U菌K1v膜血清型菌株NTUH-K2044共同培B，分x出能蚋腥敬司曛18噬菌w候x。噬菌w化、DNA萃取c限制性核酸惹忻福EcoRI、HindIIIcPstI）剪切反弥@些噬菌w均殡p股DNA噬菌w，再由其相同限制性片段L度多B性（restrictionfragmentlengthpolymorphism,RFLP）判e橄嗤删w，K依未笮☆A估其基因M大小s40Kbp。取噬菌wDNA片段序列cNCBIBLASTY料e的DNAc胺基酸序列做比Γ橐蝗挛粗删w，且凫ΦKMV噬菌w群一T。此外，使用穿透式子@微R（TEM）可^察到此噬菌w型B。透^Tc（Spottest）M行此噬菌w宿主治觯lF其Ω腥究死撞戏窝U菌K1v膜血清型菌株具有Ｒ恍浴Ｄ壳耙淹瓿伸妒删wDNA大片段定序（42.5Kbp），cM行v膜多醣降解，Y果@示v膜多醣可能降解成以熙辨Y之分子存在。在未恚耸删w⒂C渺毒N或菌株之a定，以及}合疫苗l展。
英文摘要Klebsiellapneumoniaecausedcommunity-acquiredpyogenicliverabscessisanemerginginfectiousdiseaseandisoftencomplicatedwithmetastaticendophthalmitisandsepticmeningitis.Capsularserotypes,especiallyK1andK2,werereportedtoplayanimportantroleinpathogenesisofK.pneumoniaecausingpyogenicliverabscess.Inthisstudy,bacteriophageswereisolatedfromtheco-cultureofserotypeK1K.pneumoniaeNTUH-K2044strainandsewage.Werandomlyselectedeighteenbacteriophagesforfurtherstudy.Byexperimentsofrestrictionenzymedigestion,wehaveknownthatthegenomeofbacteriophagesweredouble-strandedDNAandthelengthofrestrictionfragmentsamongtheseeighteenbacteriophageswereidentical.Thegenomesizeofbacteriophagewaspredictedtobeabout40kbp,andaroughmorphologyofbacteriophagewasobservedbytransmissionelectronmicroscope.OurpreliminaryresultrevealedthatthesequencesofthisbacteriophagewasnovelbycomparisonwithNCBIBLASTdatabases,andwehadaDNAframentcoveringabout95%areaofbacteriophagegenome.Withspottestforhostrange,wefoundthenewbacteriophagespecificallyinfectedK.pneumoniaeserotypeK1.Inaddition,it’sbeenknownthatcapsularpolysaccharidedegradedtooligomersafterphagetreatmentcouldbedetectedbyWesternblottingandmassspectrometry.Thisbacteriophagemightbeagoodcandidatefortyping,andhelpsthedevelopmentofconjugatevaccine.
文目次D目.....................................................................................................3
表目.....................................................................................................4
中文摘要.................................................................................................5
英文摘要.................................................................................................6
第一章、w........................................................................................7
第二章、材料c方法..........................................................................11
1.菌株...........................................................................................11
2.|w及培B基....................................................................................11
3.ふ壹胺蛛x噬菌w............................................................................11
4.Tc............................................................................................13
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