Dithiothreitol (DTT) chinhu chinoshandiswa zvakanyanya kuderedza, chinozivikanwawo sechinowedzerwa chitsva chegirini. Ichinhu chidiki cheorganic chemorekuru chine mapoka maviri emercaptan (-SH). Nekuda kwehunhu hwayo hwekuderedza uye kugadzikana, DTT inoshandiswa zvakanyanya mukuyedza biochemistry uye molecular biology.
Basa guru reDTT nderekuderedza zvisungo zve disulfide mumapuroteni nemamwe ma biomolecules. Chisungo che disulfide chikamu chakakosha chekupeta nekugadzikana kwemapuroteni, asi pasi pemamwe mamiriro ekuyedza, akadai sekuongorora kweSDS-PAGE kunoderedzwa, kusanganiswa kwemapuroteni nekupeta, zvakakosha kuderedza chisungo che disulfide kumapoka maviri e thiol kuti pave nekupatsanurwa kwechimiro che spatial protein. DTT inogona kuita ne disulfide bonds kuti ivaderedze kuita mapoka e mercaptan, nokudaro ichivhura chimiro che spatial protein uye zvichiita kuti zvive nyore kuongorora nekugadzirisa.
DTT inogona zvakare kushandiswa kuchengetedza mashandiro ema enzyme uye kugadzikana. Mune mamwe maitiro e enzyme-catalyzed, mashandiro ema enzyme anogona kuderedzwa ne oxidant. DTT inogona kuita nema oxidants kuti iaite zvinhu zvisina njodzi, nokudaro ichidzivirira mashandiro uye kugadzikana kwema enzyme.
Zvichienzaniswa nezvinhu zvechinyakare zvinoderedza simba zvakaita seβ-mercaptoethanol (β-ME), DTT inoonekwa senzira yekuderedza simba yakachengeteka uye yakagadzikana. Haingogadzikane mumvura chete, asiwo inochengetedza hunhu hwayo hwekuderedza simba mumamiriro ekupisa kwakanyanya uye acid-base.
Kushandisa DTT kuri nyore. Kazhinji, DTT inonyungudutswa mubuffer yakakodzera yozowedzerwa musystem yekuedza. Kuwanda kwakakwana kweDTT kunofanirwa kuongororwa zvichienderana nekuyedza kwacho, uye kunowanzo shandiswa muhuwandu hwe0.1-1mM. Kuwanda kwakaderera kunogona kuderedza mhedzisiro yakaipa pakukura kwemasero uye kunogona kuderedza kukuvadza kwemasero nekuda kwekuwedzera kwemapuroteni anotarisirwa. Kuwanda kwakawanda kunogona kukonzera mutoro wakawandisa wemetabolism yemasero, zvichikanganisa kukura kwemasero uye kushanda zvakanaka kwekutaura.
Nzira yekuona huwandu hwakakodzera inogona kuve yekuongorora huwandu hweproteni inotarwa nekuita bvunzo dzeIPTG dzekutanga pahuwandu hwakasiyana. Miedzo midiki yekumera inogona kuitwa uchishandisa huwandu hwakasiyana hweIPTG (semuenzaniso 0.1 mM, 0.5 mM, 1 mM, nezvimwewo) uye mhedzisiro yekutaura pahuwandu hwakasiyana inogona kuongororwa nekuona huwandu hweproteni inotarwa (semuenzaniso Western blot kana fluorescence detection). Zvichienderana nemhedzisiro yekuyedza, huwandu hwakanakisisa hwekutaura hwakasarudzwa sehuwandu hwakanakisisa.
Pamusoro pezvo, unogona zvakare kutarisa mabhuku akakodzera kana ruzivo rwemamwe marabhoritari kuti unzwisise huwandu hwehuwandu hweIPTG hunoshandiswa zvakanyanya pasi pemamiriro akafanana ekuyedza, wobva wagadzirisa uye wagadzirisa zvinoenderana nezvinodiwa zvekuyedza.
Zvakakosha kuziva kuti huwandu hwakakodzera hwemasero hunogona kusiyana zvichienderana nemaitiro akasiyana ekutaura, mapuroteni anotarisirwa, uye mamiriro ekuyedza, saka zvakanaka kugadzirisa mamiriro acho ezvinhu zvichienderana nemamiriro ezvinhu.
Muchidimbu, DTT chinhu chinoshandiswa zvakanyanya kuderedza ma "disulfide bonds" mumapuroteni nemamwe ma "biomolecules" uye kuchengetedza mashandiro ema "enzymes" uye kugadzikana. Chave chichishandiswa zvakanyanya mukuyedza biochemistry uye molecular biology.
Nguva yekutumira: Gunyana-28-2023