Indlela yokunquma ukuhlushwa okuphelele?
Ku-Inducer ye-IPTG (isopropyl-beta-d-thiogalactoside), ukugxila okuphezulu, kuba ngcono.Ukugxiliswa okuhle kakhulu kuncike ezimweni ezithile zokuhlola kanye nomphumela odingekayo wokungenisa.
Ngokuvamile, ukugxila kwe-IPTG kusetshenziswa kububanzi obungu-0.1-1 mM.Ukugxila okuphansi kunganciphisa imiphumela emibi ekukhuleni kweseli futhi kunganciphisa i-cytotoxicity ngenxa yokweqisa ngokweqile kwamaprotheni okuhlosiwe.Ukugxila okuphezulu kungase kubangele umthwalo omkhulu we-cell metabolism, kuthinte ukukhula kwamangqamuzana nokusebenza kahle kwenkulumo.
Indlela yokunquma ukugxilisa ingqondo okuphelele kungaba ukuhlola izinga lenkulumo yephrotheni eqondiwe ngokwenza izivivinyo zokungeniswa kwe-IPTG ezindaweni ezihlukene.Ukuhlolwa kwesiko lezinga elincane kungenziwa kusetshenziswa uhla lokugxilisa kwe-IPTG (isb. 0.1 mM, 0.5 mM, 1 mM, njll.) kanye nomthelela wenkulumo ekugxilweni okuhlukene kungahlaziywa ngokuthola izinga lenkulumo yephrotheni eqondiwe (isb. ukubona i-blot noma i-fluorescence).Ngokusho kwemiphumela yokuhlola, ukugxilisa ingqondo okunomphumela omuhle kakhulu wokuveza kukhethwe njengokugxilisa okungcono kakhulu.
Ngaphezu kwalokho, ungaphinda ubhekisele ezincwadini ezifanele noma ulwazi lwamanye amalabhorethri ukuze uqonde ububanzi bokuhlushwa be-IPTG obusetshenziswa kakhulu ngaphansi kwezimo zokuhlola ezifanayo, bese ulungiselela futhi ulungise ngokuya ngezidingo zokuhlola.
Kubalulekile ukuqaphela ukuthi ukugxilisa ingqondo okuhle kungase kwehluke kuye ngamasistimu enkulumo ahlukene, amaprotheni okuqondiwe, nezimo zokuhlola, ngakho-ke kungcono kakhulu ukukhulisa ngokwesisekelo secala ngalinye.
Isikhathi sokuthumela: Sep-28-2023