Dithiothreitol (DTT) yog ib qho tshuaj txo qis, tseem hu ua cov tshuaj ntsuab tshiab.Nws yog ib qho me me molecular organic compound nrog ob pawg mercaptan (-SH).Vim nws txo qis zog thiab ruaj khov, DTT yog dav siv hauv biochemistry thiab molecular biology thwmsim.
Lub luag haujlwm tseem ceeb ntawm DTT yog txo qis disulfide bonds hauv cov protein thiab lwm yam biomolecules.Lub disulfide daim ntawv cog lus yog ib qho tseem ceeb ntawm cov protein folding thiab stability, tab sis nyob rau hauv tej yam kev sim, xws li reducible SDS-PAGE tsom xam, protein recombination thiab folding, nws yog tsim nyog los txo cov disulfide daim ntawv cog lus rau ob pawg thiol kom unravel lub spatial qauv ntawm. cov protein.DTT tuaj yeem cuam tshuam nrog disulfide daim ntawv cog lus kom txo tau lawv mus rau mercaptan pawg, yog li qhib cov qauv spatial ntawm cov protein thiab ua kom yooj yim rau kev txheeb xyuas thiab tswj xyuas.
DTT kuj tseem siv tau los tiv thaiv kev ua haujlwm enzyme thiab kev ruaj ntseg.Hauv qee cov tshuaj tiv thaiv enzyme-catalyzed, cov haujlwm ntawm cov enzyme yuav raug txo los ntawm oxidant.DTT tuaj yeem hnov mob nrog oxidants kom txo tau lawv rau cov khoom tsis zoo, yog li tiv thaiv kev ua haujlwm thiab kev ruaj ntseg ntawm cov enzyme.
Piv nrog rau cov tshuaj txo cov tshuaj xws li β-mercaptoethanol (β-ME), DTT raug suav tias yog tus neeg saib xyuas kev nyab xeeb thiab ruaj khov dua.Nws tsis tsuas yog ruaj khov hauv cov kua dej, tab sis kuj tswj nws cov khoom txo qis hauv qhov kub thiab txias thiab acid-puag puag.
Kev siv DTT yog qhov yooj yim.Feem ntau, DTT yog yaj nyob rau hauv ib qho tsis haum thiab tom qab ntawd ntxiv rau qhov kev sim.Qhov kev pom zoo ntawm DTT yuav tsum tau txiav txim siab raws li qhov kev sim tshwj xeeb, thiab feem ntau yog siv nyob rau hauv thaj tsam ntawm 0.1-1mM.Cov concentrations qis dua tuaj yeem txo qhov tsis zoo ntawm kev loj hlob ntawm tes thiab tuaj yeem txo cytotoxicity vim overexpression ntawm lub hom phiaj proteins.Cov concentration ntau dua tuaj yeem ua rau lub nra hnyav ntawm cov cell metabolic, cuam tshuam rau kev loj hlob ntawm tes thiab kev nthuav qhia kev ua haujlwm.
Txoj hauv kev los txiav txim siab qhov zoo tshaj plaws concentration tuaj yeem ntsuas qhov qhia theem ntawm lub hom phiaj protein los ntawm kev ua IPTG induction xeem ntawm ntau qhov sib txawv.Kev kuaj kab lis kev cai me me tuaj yeem ua tau siv ntau yam ntawm IPTG concentrations (piv txwv li 0.1 mM, 0.5 mM, 1 mM, thiab lwm yam) thiab cov txiaj ntsig ntawm qhov sib txawv tuaj yeem ntsuas los ntawm kev txheeb xyuas qhov qhia theem ntawm lub hom phiaj protein (xws li Western blot los yog fluorescence detection).Raws li kev sim cov txiaj ntsig, qhov concentration nrog cov txiaj ntsig zoo tshaj plaws tau xaiv los ua qhov kev pom zoo.
Tsis tas li ntawd, koj tuaj yeem xa mus rau cov ntaub ntawv ntsig txog lossis kev paub ntawm lwm lub chaw kuaj mob kom nkag siab txog qhov feem ntau siv IPTG concentration ntau nyob rau hauv cov kev sim zoo sib xws, thiab tom qab ntawd kho kom zoo thiab kho raws li qhov xav tau ntawm kev sim.
Nws yog ib qho tseem ceeb uas yuav tsum nco ntsoov tias qhov zoo tshaj plaws concentration yuav txawv nyob ntawm seb cov kab lus sib txawv, lub hom phiaj ntawm cov proteins, thiab cov kev sim, yog li nws yog qhov zoo tshaj plaws los ua kom zoo dua ntawm rooj plaub.
Nyob rau hauv cov ntsiab lus, DTT yog ib tug nquag siv txo tus neeg sawv cev uas yuav siv tau los txo disulfide bonds nyob rau hauv cov proteins thiab lwm yam biomolecules thiab los tiv thaiv enzyme kev ua si thiab stability.Nws tau siv dav hauv biochemistry thiab molecular biology thwmsim.
Post lub sij hawm: Sep-28-2023