Dithiothreitol (DTT) nikintu gikunze kugabanya kugabanya, kizwi kandi nk'icyatsi gishya.Nibintu bito bya molekuline kama hamwe nitsinda rya mercaptan ebyiri (-SH).Bitewe no kugabanya imiterere no gutuza, DTT ikoreshwa cyane mubinyabuzima na bioologiya ya biologiya.
Uruhare nyamukuru rwa DTT ni ukugabanya imiyoboro ya disulfide muri poroteyine no mu bindi binyabuzima.Inkunga ya disulfide nigice cyingenzi cyububiko bwa poroteyine no gutuza, ariko mubihe bimwe byubushakashatsi, nko kugabanya isesengura rya SDS-PAGE, isesengura rya poroteyine no kuzunguruka, birakenewe kugabanya umurunga wa disulfide mumatsinda abiri ya thiol kugirango uhishure imiterere yumwanya wa poroteyine.DTT irashobora kwitwara hamwe na disulfide kugirango igabanye mumatsinda ya mercaptan, bityo ifungura imiterere yimiterere ya poroteyine kandi byoroshye kuyisesengura no kuyikoresha.
DTT irashobora kandi gukoreshwa mukurinda ibikorwa bya enzyme no gutuza.Mubisubizo bimwe na bimwe bya enzyme-catalizike, ibikorwa bya enzyme birashobora kugabanuka na okiside.DTT irashobora kwitwara hamwe na okiside kugirango igabanye ibintu bitagira ingaruka, bityo irinde ibikorwa nibitekerezo bya enzyme.
Ugereranije no kugabanya imiti gakondo nka β-mercaptoethanol (β-ME), DTT ifatwa nkigikoresho cyizewe kandi gihamye cyo kugabanya.Ntabwo ihagaze neza mugisubizo cyamazi gusa, ahubwo inagumana imiterere yayo igabanya ubushyuhe bwinshi hamwe na aside-ishingiro.
Gukoresha DTT biroroshye.Muri rusange, DTT iseswa muri buffer ikwiye hanyuma ikongerwaho sisitemu yubushakashatsi.Ibyiza bya DTT bigomba kugenwa ukurikije igeragezwa ryihariye, kandi muri rusange bikoreshwa murwego rwa 0.1-1mM.Kwibanda cyane birashobora kugabanya ingaruka mbi kumikurire ya selile kandi birashobora kugabanya cytotoxicité bitewe no gukabya gukabije kwa poroteyine.Kwibanda cyane birashobora gutera uburemere bwimikorere ya selile, bikagira ingaruka kumikurire no gukora neza.
Inzira yo kumenya icyerekezo cyiza irashobora kuba ugusuzuma urwego rwimiterere ya poroteyine igamije gukora ibizamini bya induction ya IPTG mubitekerezo bitandukanye.Ibizamini byumuco muto birashobora gukorwa hifashishijwe urutonde rwa IPTG (urugero: 0.1 mM, 0.5 mM, 1 mM, nibindi) kandi ingaruka zigaragaza mubitekerezo bitandukanye zishobora gusuzumwa no kumenya urwego rwerekana poroteyine (urugero: Iburengerazuba blot cyangwa fluorescence detection).Ukurikije ibisubizo byubushakashatsi, kwibanda hamwe ningaruka nziza zerekana imvugo byatoranijwe nkibintu byiza.
Wongeyeho, urashobora kandi kwifashisha ibitabo bijyanye cyangwa uburambe bwizindi laboratoire kugirango usobanukirwe na IPTG ikunze gukoreshwa mugihe cyibigeragezo bisa, hanyuma ugahindura kandi ugahindura ukurikije ibikenewe mubushakashatsi.
Ni ngombwa kumenya ko kwibandaho kwiza gushobora gutandukana bitewe na sisitemu zitandukanye zerekana imvugo, intego za poroteyine, hamwe nubushakashatsi bwakozwe, nibyiza rero guhitamo neza kuri buri kibazo.
Muncamake, DTT nikintu gikoreshwa mukugabanya kugabanya bishobora gukoreshwa mukugabanya imiyoboro ya disulfide muri proteyine nizindi biomolecules no kurinda ibikorwa bya enzyme no gutuza.Yakoreshejwe cyane mubinyabuzima na molekuline ya biologiya.
Igihe cyo kohereza: Nzeri-28-2023