Dithiothreitol (DTT) ni ikintu gikoreshwa cyane mu kugabanya ibintu, kizwi kandi nk'inyongera nshya y'icyatsi kibisi. Ni ikintu gito cy'umwimerere gifite amatsinda abiri ya mercaptan (-SH). Bitewe n'imiterere yacyo igabanya ibintu kandi ikagumana ubuziranenge, DTT ikoreshwa cyane mu bushakashatsi bwa biochimie na molekile.
Inshingano nyamukuru ya DTT ni ukugabanya imigozi ya disulfide muri poroteyine n'izindi biomolecules. Imigozi ya disulfide ni igice cy'ingenzi mu gupfunyika no guhagarara kwa poroteyine, ariko mu bihe bimwe na bimwe by'igerageza, nko gusesengura SDS-PAGE igabanuka, gusubiranamo kwa poroteyine no gupfunyika, ni ngombwa kugabanya imigozi ya disulfide ku matsinda abiri ya thiol kugira ngo haveho imiterere y'ahantu ya poroteyine. DTT ishobora gukorana n'imigozi ya disulfide kugira ngo igabanuke ku matsinda ya mercaptan, bityo igafungura imiterere y'ahantu ya poroteyine kandi bigatuma byoroha kuyisesengura no kuyihindura.
DTT ishobora kandi gukoreshwa mu kurinda imikorere ya enzyme no kudahinduka. Mu buryo bumwe na bumwe butuma enzyme ikora, imikorere ya enzyme ishobora kugabanuka bitewe na oxidant. DTT ishobora gukorana na oxidant kugira ngo igabanukemo ibintu bitagize ingaruka mbi, bityo ikarinda imikorere n'uburyo enzyme ikora.
Ugereranyije n'ibintu bisanzwe bigabanya ubushyuhe nka β-mercaptoethanol (β-ME), DTT ifatwa nk'ikintu gifasha mu kugabanya ubushyuhe kandi kidahindagurika. Ntabwo ihoraho gusa mu mazi, ahubwo inakomeza ubushobozi bwayo bwo kugabanya ubushyuhe mu gihe cy'ubushyuhe bwinshi n'imiterere y'aside.
Gukoresha DTT biroroshye cyane. Muri rusange, DTT ishongeshwa mu cyuma gikwiye hanyuma ikajya mu buryo bw'igerageza. Igipimo cyiza cya DTT kigomba kugenwa hakurikijwe igerageza runaka, kandi muri rusange gikoreshwa hagati ya 0.1-1mM. Igipimo gito gishobora kugabanya ingaruka mbi ku mikurire y'uturemangingo kandi gishobora kugabanya uburozi buterwa no kwiyongera cyane kwa poroteyine. Igipimo kinini gishobora gutera umutwaro ukabije w'imikorere y'uturemangingo, bigagira ingaruka ku mikurire y'uturemangingo no ku mikorere myiza yatwo.
Uburyo bwo kumenya urugero rwiza rushobora kuba ugusuzuma urugero rw'igaragara rya poroteyine y'intego hakoreshejwe ibizamini bya IPTG ku bipimo bitandukanye. Ibizamini bito byo gupima bishobora gukorwa hakoreshejwe ibipimo bitandukanye bya IPTG (urugero: 0.1 mM, 0.5 mM, 1 mM, nibindi) kandi ingaruka z'igaragara ku bipimo bitandukanye zishobora gusuzumwa hakoreshejwe kumenya urugero rw'igaragara rya poroteyine y'intego (urugero: Western blot cyangwa fluorescence detection). Dukurikije ibyavuye mu bushakashatsi, urugero rufite ingaruka nziza z'igaragara rwatoranyijwe nk'urugero rwiza.
Byongeye kandi, ushobora kandi kureba inyandiko zijyanye n'ibyo cyangwa ubunararibonye bw'izindi laboratwari kugira ngo wumve urwego rw'ibipimo bya IPTG bikunze gukoreshwa mu bihe bisa byo kugerageza, hanyuma urusheho kunoza no guhindura uko ubyifuza mu igerageza.
Ni ngombwa kumenya ko ingano nziza ishobora gutandukana bitewe n'uburyo butandukanye bwo kugaragaza, poroteyine zigamije, n'imiterere y'igerageza, bityo ni byiza kunoza buri kimwe uko cyakabaye.
Muri make, DTT ni ikintu gikoreshwa cyane mu kugabanya imigozi ya disulfide muri poroteyine n'izindi biomolecules no kurinda imikorere ya enzymes no guhagarara kwayo. Yakoreshejwe cyane mu bushakashatsi bwa biochimie na molekile.
Igihe cyo kohereza ubutumwa: 28 Nzeri 2023