Dithiothreitol (DTT) inowanzo shandiswa kuderedza mumiriri, inozivikanwawo seyakasvibira yekuwedzera.Idiki mamolecular organic compound ine maviri mercaptan mapoka (-SH).Nekuda kwekudzikisa kwayo zvivakwa uye kugadzikana, DTT inoshandiswa zvakanyanya mubiochemistry uye mamorekuru biology kuyedza.
Basa guru reDTT ndere kuderedza disulfide zvisungo mumapuroteni uye mamwe mabiomolecules.Iyo disulfide bond chikamu chakakosha cheprotein kupeta uye kugadzikana, asi pasi pemamwe mamiriro ekuyedza, akadai seReducible SDS-PAGE kuongororwa, protein recombination uye kupeta, zvinodikanwa kuderedza disulfide bond kumapoka maviri etiol kuti aburitse chimiro chepakati iyo protein.DTT inogona kuita ne disulfide zvisungo kuti zvideredze kumapoka e mercaptan, nokudaro kuvhura chimiro chepakati cheprotein uye kuita kuti zvive nyore kuongorora nekushandisa.
DTT inogona zvakare kushandiswa kuchengetedza enzyme basa uye kugadzikana.Mune mamwe maitiro e enzyme-catalyzed, basa re enzyme rinogona kuderedzwa neiyo oxidant.DTT inogona kuita nemaoxidants kuti ivaderedze kuzvinhu zvisingakuvadzi, nekudaro kuchengetedza chiitiko uye kugadzikana kweiyo enzyme.
Kuenzaniswa nemaitiro ekudzikisa echinyakare akadai se β-mercaptoethanol (β-ME), DTT inoonekwa seyakachengeteka uye yakagadzikana kuderedza mumiriri.Haisi kungogadzikana mukugadzirisa kwemvura, asiwo inochengetedza maitiro ayo ekuderedza pasi pekushisa kwepamusoro uye acid-base mamiriro.
Kushandiswa kweDTT kuri nyore.Kazhinji, DTT inonyungudutswa mune yakakodzera buffer uye yozowedzerwa kune yekuyedza system.Iyo yakakwana yekusangana kweDTT inoda kutariswa zvichienderana nekuyedza chaiyo, uye inowanzoshandiswa muhuwandu hwe0.1-1mM.Kudzika kwakadzika kunogona kuderedza mhedzisiro yakashata pakukura kwesero uye kunogona kuderedza cytotoxicity nekuda kwekuwedzeredza kwechinangwa cheprotein.Kukwirira kwakanyanya kunogona kukonzera kuwandisa kwesero metabolic mutoro, zvichikanganisa kukura kwesero uye kugona kutaura.
Iyo nzira yekuona iyo yakakwana yekuisa inogona kuve yekuongorora iyo yekutaura nhanho yeinonangwa puroteni nekuita IPTG induction bvunzo pane dzakasiyana siyana.Zviyedzo zvetsika diki-diki zvinogona kuitwa pachishandiswa huwandu hwe IPTG (eg 0.1 mM, 0.5 mM, 1 mM, zvichingodaro) uye mhedzisiro yekutaura pane zvakasiyana-siyana inogona kuongororwa nekuona danho rekutaura reprotein yakanangwa (semuenzaniso Western. blot kana fluorescence kuonekwa).Zvinoenderana nemhedzisiro yekuyedza, iyo yekumisikidza ine yakanakisa kutaura mhedzisiro yakasarudzwa seyakanyanya kugadzirisa.
Pamusoro pezvo, iwe unogona zvakare kutarisa kune akakodzera mabhuku kana ruzivo rwemamwe marabhoritari kuti unzwisise inowanzoshandiswa IPTG yekumisikidza renji pasi pemamiriro ekuyedza akafanana, wobva wawedzera uye kugadzirisa zvinoenderana nezvinodiwa zvekuedza.
Izvo zvakakosha kuti uzive kuti iyo optimal concentration inogona kusiyana zvichienderana neakasiyana ekutaura masisitimu, anonangwa mapuroteni, uye kuyedza mamiriro, saka zviri nani kukwirisa pane imwe-ne-kesi hwaro.
Muchidimbu, DTT inowanzoshandiswa kuderedza mumiririri inogona kushandiswa kuderedza disulfide zvisungo mumapuroteni uye mamwe biomolecules uye kuchengetedza enzyme basa uye kugadzikana.Yakave ichishandiswa zvakanyanya mubiochemistry uye molecular biology kuyedza.
Nguva yekutumira: Sep-28-2023