Indlela yokumisela ugxininiso olufanelekileyo?
Kwi-inducer IPTG (isopropyl-beta-d-thiogalactoside), iphezulu i-concentration, ingcono.Ugxininiso olufanelekileyo luxhomekeke kwiimeko ezithile zovavanyo kunye nesiphumo esifunwayo sokungeniswa.
Ngokuqhelekileyo, ukuxinwa kwe-IPTG kusetyenziswe kuluhlu lwe-0.1-1 mM.Ukugxininiswa okuphantsi kunokunciphisa imiphumo emibi ekukhuleni kweeseli kwaye kunokunciphisa i-cytotoxicity ngenxa yokugqithiswa kweeprotheni ezijoliswe kuyo.Ukugxila okuphezulu kunokubangela umthwalo ogqithisileyo we-cell metabolic, echaphazela ukukhula kweeseli kunye nokusebenza kakuhle kokubonakalisa.
Indlela yokumisela ugxininiso olufanelekileyo lunokuvavanya inqanaba lokuvakalisa iprotheni ekujoliswe kuyo ngokuqhuba iimvavanyo ze-IPTG zokungeniswa kwiindawo ezahlukeneyo.Uvavanyo lwenkcubeko olukwizinga eliphantsi lunokwenziwa kusetyenziswa uluhlu lwe-IPTG yogxininiso (umz. 0.1 mM, 0.5 mM, 1 mM, njl. njl.) kunye nesiphumo sokubonakaliswa kwiimpawu ezahlukeneyo sinokuvavanywa ngokufumanisa inqanaba lokubonisa iprotheni ekujoliswe kuyo (umz. ukufunyanwa kwebala okanye ukubonwa kwe-fluorescence).Ngokweziphumo zovavanyo, ugxininiso olunesiphumo esihle sokubonisa sikhethwe njengogxininiso olufanelekileyo.
Ukongeza, unokubhekisa kuncwadi olufanelekileyo okanye amava ezinye iilabhoratri ukuqonda uluhlu loxinaniso lwe-IPTG olusetyenziswa ngokuqhelekileyo phantsi kweemeko zovavanyo ezifanayo, kwaye emva koko ulungise kwaye ulungelelanise ngokweemfuno zovavanyo.
Kubalulekile ukuqaphela ukuba ugxininiso olufanelekileyo lunokwahluka ngokuxhomekeke kwiinkqubo zokuvakalisa ezahlukeneyo, iiprotheyini ezijoliswe kuzo, kunye neemeko zovavanyo, ngoko ke kungcono ukukhulisa kwimeko-nge-case basis.
Ixesha lokuposa: Sep-28-2023