基于耐辐射球菌的生物传感器的构建以及耐辐射球菌drasnf2基因的研究

《基于耐辐射球菌的生物传感器的构建以及耐辐射球菌drasnf2基因的研究》由会员上传分享，免费在线阅读，更多相关内容在学术论文-天天文库。

1、突变株，经过DNA损伤实验，发现draSNF2对耐辐射球菌的极端抗辐射特性并不是必需的。VllAbstractBasedonageneticallymodifiedradioresistantbacteriaDeinococcusradiodurans，weconstructedarealtimewholecellbiosensortomonitorradioactivityandgenotoxicityinhi曲Iyradioactiveenvironment。Theerahancedgreenfluorescenceprotein(eGFP)wasfusedtothepromoter

2、ofacrucialDNAdamage—induciblerecAgenefrom／9．radiodurans，andtheconsequentDNAfragment(PrecA-eg兹o)carriedbyplasmidwasintroducedintonradioduransR1straintoobtainthebiosensorstrainDRG300．ThisengineeredstrainCallexpresseGFPproteinandgeneratefluorescenceininductionoftherecAgenepromoter．Basedonthecorrelat

3、ionbetweenfluorescenceintensityandproteinexpressionlevelinliveD：radioduranscells，wediscoveredthatthefluorescenceinductionofstrainDRG300respondsinaremarkabledose-dependentmannerwhentreated诚m转NAdamagesourcessuchasgammaradiationandmitomycinC。Itisencouragingtofindthewidelydetectiverangeandhi曲sensitiv

4、ityofthisreconstructedstraincomparing、蕊也otherwholecellbiosensorsinformerreports．TheseresultssuggestthestrainDRG300apotentialwholecellbiosensortoconstructadetectivesystemtomonitorthebiolo西calhazardsofradioactiveandtoxicpollutantsinenvironmentinrealtime．弧eBeta-Glo癣AssaySystemisahomogeneousmethodofq

5、uantitating13-galactosidaseexpressioninmammaliancells+强esystemprovidesabrightluminescentsignalthatisstableoverseveralhoursincommonlyusedcellculturemediumwithoutpriorsampleprocessing．Thehomogeneousassayprocedureinvolvestheadditionofasinglereagentdirectlytocellsculturedinserum-supplementedmedium。Th

6、roughputratesofseveralthousandsamplesperhourmaybeachieved谢也hi曲reproducibilityunderstandardlaboratoryconditions．WiththisIleWtechnology，weconstructedanewsystemforwholeeellbiosensor．HomologsofSnf2familymemberin叠radioduransR1，draSNF2WassequencingedtodefineasingleORFincludingtheon-linetagDRl258andDRl2

7、59。ThenewdefineddraSNF2genewithouttheinteinsequenceWasanalyzedwithSSO1653，andpredicteditstructureandbiologicalfunction．StudybytreatingDradioduransRlstrainsanddraSNF2function—deficientmutantwithionizingradiationandMMCwe