噬菌體展示免疫球蛋白結(jié)合分子單結(jié)構(gòu)域隨機組合文庫的構(gòu)建及體外分子進化.pdf

1、山西醫(yī)科大學碩士學位論文噬菌體展示免疫球蛋白結(jié)合分子單結(jié)構(gòu)域隨機組合文庫的構(gòu)建及體外分子進化姓名：楊華申請學位級別：碩士專業(yè)：微生物學指導教師：李連青20050520山西最辯大學磺士學位論文Abstractobjective：Throughcomparingthecourseofmoleculeevolutionalselectioninvitroofthephage—displayedrandomcombinatoriallibrar

2、yofsingledomainsofIg—bindingmoleculesleadedbydifferentIgmoleculesandthestmctureandfunctionoftherecombinantIgbindingmoleculesobtainedbytheselection，WeaimedtoexploretherelationshipbetweenstructureandfunctionofbacteriaIgbin

3、dingmolecules，themechanismoftheinteractionbetweenthemandIgmolecules，andtherulesoftheirrecombinantandevolutioninvitroinordertoaccumulateexpedencesofreconstructionoffunctionalproteinsusingthemethodsofmolecularevolutioninfu

4、tureMethods：ThemoleculardirectedevolutionaiaffinityselectionsinvitroofaphagedisplayedrandomcombinatoriallibraryoftheA，DdomainsofproteinA，theB2domainsofproteinQtheB3domainsofproteinLlinkedbytherandomlinkingpeptideconsiste

5、dof0，1，2or3aminoacidswereprocessedabout34timesusingindividuallyhumanIgGIgM，]gAantibodymoleculesandIgGFcconstructedbygeneengineeringaspanningtargetsWeselectedeightrepresentativedisplayedsequenceshavingdifferentstructurefo

6、rmswhichwereobtainedbytheinducementsofdifferentlgmoleculesandclonedthemtotheprokaryoticexpressionvectorPET32a()，thenexpressedthefusionproteinswithHisTagontheNterminal，whichwerepurifiedwimNi_NTAresin，andcharacterizedtheir

7、IgbindingactivitybyWesternblotandELISAResult：ThecombinatoriallibraryshowedsimilarchangeduringtheprocessofaffinityselectionsleadedbyIgMandIgAandthedisplayedsequenceswereevolvedintoacommonandnewstructureform：PAA—PLwhichdoe

8、sn’texistinbacteriaIgbindingmoleculesinnatureWehavegotsixdifferentsequencesofthisformbecauseofthedifferentsequencesoflinldngpeptides，butthesesequencestrendedtoappearintheevolutionlibrariesofIgMand]gAatthesanletimeTheevol