18788.干擾素調(diào)節(jié)因子7dna結(jié)合結(jié)構(gòu)域中賴氨酸殘基的作用

1、南開大學碩士學位論文干擾素調(diào)節(jié)因子7DNA結(jié)合結(jié)構(gòu)域中賴氨酸殘基的作用姓名：張敏申請學位級別：碩士專業(yè)：微生物學指導教師：王金忠201205AbstractInterferonregulatoryfactor7(IRF7)isamemberoftheIRFfamilywhichplaysimportantrolesintwoaspectsOneisregulatingtheexpressionofinterferonandtheothe

2、risDarticipatingintheantiviralimmuneresponseIRF一7isaDNAbindingproteincontaining503aminoacidsTheN—terminal151aminoacidresiduesofIRF7constituteitsDNAbindingdomain(DBD)，whichisresponsibleforitsbindingtotheinterferonstimulat

3、edresponseelement(ISRE)ThefunctionofIRF7carlberegulatedbyposttranslationalmodificationsincludingphosphorylation，acetylation，ubiquitinationandSUMOylationLysineresiduesarepotentialsitestbrthesemodifications，andmayserveasta

4、rgetstomodulatethefunctionsofIRF7TherearefivelysineresiduesintheIRF7DBDIthasbeenreportedthatlysineresiduemutationSmayhaveaffectedthefunctionofIRF7，andsomelysineresiduemutaftonsintheIRF7DBDmayhaveabolishedthefunctionofIRF

5、7In“sstudv’eachofthelysineresiduesofIRF一7DBDwasmodifiedintothearginineresiduebysitedirectedmutagenesisSixexpressionplasmidswereconstmcted，whichexpressedrecombinantwildtypeorgRoRmutantIRF7DBDDroteinseachfusedwitha6xhistid

6、inetagandaFLAGtagThe6xhistidinetagandtheFLAGtagwereusedforproteinpurificationanddetectil3n，respectivelyThentherecombinantwildtypeormutantIRF一7DBDproteinswerepurified，andtheirabilitiestobindwiththeISREweredetectedusingthe

7、DNABINDmicroplateItisshownthatthewildtypeIRF7DBDcarlbindwiththeISREspecificallyComparedwimthewildtype，thebindingabilitiesofmutantsK45R，K50R，K61R，K92RandK120RwereonlV13％，54％，22％，28％and16％ofthatforthewildtype，indicatingtha

8、ttheselysineresiduemutationsaffectedthebindingabilityofIRF7DBDwiththeISRETheseresultsdemonstratedtheimportantrolethatIRF7lysineresiduesplayedinitsDNAbindingabilityandprovidedevidenceforthefurtherusageottheDNA—BINDmicropl