Yuav ua li cas txiav txim siab qhov concentration zoo tshaj plaws?
Rau tus inducer IPTG (isopropyl-beta-d-thiogalactoside), qhov concentration siab dua, qhov zoo dua. Qhov concentration zoo tshaj plaws nyob ntawm cov xwm txheej kev sim tshwj xeeb thiab cov nyhuv induction xav tau.
Feem ntau, qhov concentration ntawm IPTG yog 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 yog kev overexpression ntawm cov protein target. Cov concentration siab dua tuaj yeem ua rau muaj kev cuam tshuam ntawm cov cell metabolic 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.
Lub sijhawm tshaj tawm: Cuaj hlis-28-2023