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  • 简介:ToconstructandexpressthefusionproteinStx2B-IntiminC300ofEHEC0157:H7,andtofurtherinvestigateitsimmunoprophyiacticpotential,thegeneofStx2B(stx2b)fromEHEC0157:H7chromosomewasclonedintopMD18-Tvector.Thereafter,theamplifiedgenewasclonedintoprokaryoticexpressionplasmidpET-28a(+)-eaeC300,whichwasconstructedpreviously.TherecombinantpasmidpET-28a(+)-stx2b-eaeC300wastransformedintoE.coliBL21(DE3).Afterinducement,theproteinStx2B-IntiminC300wassuccessfullyexpressedandanalyzedwithsodiumdodecylsulfatepolyacrylamidegelelectrophoresis(SDS-PAGE),WesternblottingandN-terminalaminoacidresidualsequencing.Toevaluateitsimmunoprophyiacticpotential,itwasprimarilypurifiedbyion-exchangechromatographyandinjectedinto30BALB/cmicewithAl(OH)3inthesubscapularregion.Tendaysafterthelastboostervaccination,20micewereattackedwithEHEC0157:H7lysateandtheprotectiveefficacywasobserved.Inthepresentstudy,thegeneofStx2B-IntiminC300wassuccessfullyclonedintopET-28a(+)vector.TheresultsofSDS-PAGEandWesternblottingassayshowedthatthefusionproteinwassuccessfullyexpressedintheinclusionbodyform,accountingfor25%oftotalexpressionproducts,anditsmolecularweightwasabout43kDa.TheresultoftheN-terminalaminoacidresidualsequencingshowedthatitwasidenticaltothatofthemoleculardesigned.Thepuritywasabout75%afterprimarypurification.AnimaltestsrevealedthatthefusionproteinStx2B-IntiminC300haselicitedhightiterofprotectiveantibodyrelatively.TheseresultsdemonstratethatthefusionproteinStx2B-IntiminC300issuccessfullyexpressedinprokaryoticexpressionsystemandshowscertainimmunoprophyiacticpotential.

  • 标签: 疫苗 免疫机制 蛋白质 原核表达
  • 简介:【摘要】:宫颈癌是女性生殖系统最常见的恶性肿瘤,严重危害女性的身心健康。现已明确[1]人乳头状瘤病毒(human papillomavirus,HPV)持续感染是引发女性宫颈癌发病的主要原因,其中HPV E6/E7蛋白的持续表达在宫颈癌发生发展过程中起重要作用,本文对 HPV E6/E7mRNA 的研究进展进行综述,来探讨人乳头状瘤病毒(HPV)E6/E7mRNA 检测在宫颈病变研究中的价值。

  • 标签: 人乳头状瘤病毒 E6E7mRNA 宫颈病变 综述
  • 简介:ThegenomeoftheenterohemorrhagicEscherichiacoliO157:H7EDL933contains177“O”-islands(OIs).TostudytheirpotentialcontributiontotheO157-specificpathogenicity,wesurveyedthedistributionof22OIsbyPCRandDNAhybridizationin17isolatesofShigatoxinproducing(Stx-positive)E.coliO157:H7,andcomparedwiththeirdistributionin21isolatesofStx-negativeE.coliO157and21isolatesofnon-O157entericpathogens.Fourteenof22OIswerepresentinnon-O157entericpathogensanalyzed.Eightof22OIswerefoundonlyinthe17Shigatoxin-(Stx)positiveE.coliO157:H7isolates,buttheywereabsentfromthe21Stx-negativeE.coliO157:NMandO157Hundisolatestested.Amongthe8OIs,onlyOI43orOI48wereexclusivelydetectedinStx-positiveE.coliO157:H7,absentfromneitherofStx-negativeE.coliO157andnon-O157entericpathogens,suchasSalmonella,ShigeUa,Citrobacter,Vibriocholera,enteropathogen-icE.coli(EPEC),enteroadherentE.coli(EAEC),enteroinvasiveE.coli(E1EC)andenterotoxingenicE.coli(ETEC).TheOI43andOI48are83kbinsizeandidenticalinDNAsequences,whichencodegenesforurease,telluriteresistanceandadherence.ByanalyzingtheirjunctiongeneswithPCRandDNAhybridization,wefoundthat21ChineseisolateshaveOI48only.However,for7Japanesepatientisolates,4haveOI43and3haveOI48;forAmericanisolates,2havebothofO143andOI48,2haveOI48only.ThesedataconfirmedthehighlyplasticityofthepathogenicE.coligenome.TheuniquepresenceofOI43/OI48inStx-positiveE.coli0157:H7denotesitscriticalroleinthepathogenicityspecifictothispathogen.

  • 标签: O-岛 埃希氏菌属 大肠菌O157 肠病原体 E.大肠菌 O157:H7
  • 简介:摘要:目的:探究患者对“4+7”带量采购政策认知度与认可度的调查分析。方法:以湖南省某三甲医院为例,对2020年3月至2021年3月期间到我院门诊接受诊断或治疗的1463例患者作为实验研究对象,通过调查问卷的方式对其进行调查,了解患者对“4+7”带量采购政策的认知及认可情况。结果:通过问卷调查分析来看,442例患者了解该政策,713例不太了解,其余308例患者不了解,占比分别为30.21%、48.73%、21.05%。其次,对了解、不太了解该政策的患者进行了有关的认可度调查,大部分患者认可度比较高。结论:大部分患者不太了解“4+7”带量采购政策,且了解该政策的患者对其认可度有相对较低。

  • 标签: “4+7”带量采购政策 认知度 认可度 建议