Dithiothreitol (DTT) yog ib qho tshuaj uas siv ntau heev, tseem hu ua cov tshuaj ntsuab tshiab. Nws yog ib qho tshuaj me me uas muaj ob pawg mercaptan (-SH). Vim nws cov khoom txo qis thiab kev ruaj khov, DTT siv dav hauv kev sim biochemistry thiab molecular biology.
Lub luag haujlwm tseem ceeb ntawm DTT yog txo cov disulfide bonds hauv cov protein thiab lwm yam biomolecules. Cov disulfide bond yog ib feem tseem ceeb ntawm kev sib sau ua ke thiab kev ruaj khov ntawm cov protein, tab sis nyob rau hauv qee qhov kev sim, xws li kev tshuaj xyuas SDS-PAGE uas txo tau, kev sib sau ua ke ntawm cov protein thiab kev sib sau ua ke, nws yog qhov tsim nyog los txo cov disulfide bond rau ob pawg thiol kom tshem tawm cov qauv ntawm cov protein. DTT tuaj yeem cuam tshuam nrog cov disulfide bonds kom txo lawv mus rau cov pawg mercaptan, yog li qhib cov qauv ntawm cov protein thiab ua rau nws yooj yim rau kev tshuaj xyuas thiab tswj hwm.
DTT kuj tseem siv tau los tiv thaiv kev ua haujlwm thiab kev ruaj khov ntawm cov enzyme. Hauv qee qhov kev cuam tshuam enzyme-catalyzed, qhov kev ua haujlwm ntawm cov enzyme yuav raug txo los ntawm cov oxidant. DTT tuaj yeem ua rau cov oxidants ua rau lawv txo qis rau cov tshuaj tsis muaj teeb meem, yog li tiv thaiv kev ua haujlwm thiab kev ruaj khov ntawm cov enzyme.
Piv rau cov tshuaj txo qis ib txwm muaj xws li β-mercaptoethanol (β-ME), DTT suav hais tias yog ib qho tshuaj txo qis uas muaj kev nyab xeeb dua thiab ruaj khov dua. Nws tsis tsuas yog ruaj khov hauv cov kua dej xwb, tab sis kuj tseem tswj nws cov khoom txo qis hauv qhov kub thiab txias thiab cov kua qaub-base.
Kev siv DTT yooj yim heev. Feem ntau, DTT raug yaj rau hauv ib qho buffer uas tsim nyog thiab tom qab ntawd ntxiv rau hauv qhov system sim. Qhov concentration zoo tshaj plaws ntawm DTT yuav tsum tau txiav txim siab raws li qhov kev sim tshwj xeeb, thiab feem ntau siv rau hauv qhov ntau ntawm 0.1-1 mM. Cov concentration qis dua tuaj yeem txo cov teebmeem tsis zoo rau kev loj hlob ntawm cov cell thiab tuaj yeem txo cov cytotoxicity vim muaj kev tshaj tawm ntau dhau ntawm cov protein. Cov concentration siab dua tuaj yeem ua rau muaj kev cuam tshuam ntawm cov cell metabolism ntau dhau, cuam tshuam rau kev loj hlob ntawm cell thiab kev ua haujlwm ntawm kev qhia tawm.
Txoj kev los txiav txim siab qhov concentration zoo tshaj plaws yog los ntsuam xyuas qib kev qhia tawm ntawm cov protein uas koj xav tau los ntawm kev ua qhov kev sim IPTG induction ntawm ntau yam concentration sib txawv. Kev sim kab lis kev cai me me tuaj yeem ua tiav los ntawm kev siv ntau yam IPTG concentration (piv txwv li 0.1 mM, 0.5 mM, 1 mM, thiab lwm yam) thiab qhov cuam tshuam ntawm kev qhia tawm ntawm ntau yam concentration tuaj yeem ntsuas tau los ntawm kev ntes qib kev qhia tawm ntawm cov protein uas koj xav tau (piv txwv li Western blot lossis fluorescence detection). Raws li cov txiaj ntsig ntawm kev sim, qhov concentration nrog qhov cuam tshuam zoo tshaj plaws tau raug xaiv ua qhov concentration zoo tshaj plaws.
Tsis tas li ntawd xwb, koj tuaj yeem xa mus rau cov ntaub ntawv sau txog lossis kev paub ntawm lwm lub chaw soj nstuam kom nkag siab txog qhov ntau ntawm IPTG uas siv ntau hauv cov xwm txheej sim zoo sib xws, thiab tom qab ntawd ua kom zoo dua thiab kho raws li qhov xav tau ntawm kev sim.
Nws yog ib qho tseem ceeb uas yuav tsum nco ntsoov tias qhov kev sib xyaw ua ke zoo tshaj plaws yuav txawv nyob ntawm ntau lub tshuab qhia tawm, cov protein uas lub hom phiaj, thiab cov xwm txheej sim, yog li nws yog qhov zoo tshaj plaws los ua kom zoo dua ntawm ib rooj plaub rau ib rooj plaub.
Hauv kev xaus lus, DTT yog ib qho tshuaj uas siv ntau heev uas siv tau los txo cov disulfide bonds hauv cov protein thiab lwm yam biomolecules thiab los tiv thaiv kev ua haujlwm thiab kev ruaj khov ntawm enzyme. Nws tau siv dav hauv kev sim biochemistry thiab molecular biology.
Lub sijhawm tshaj tawm: Cuaj hlis-28-2023