Kodi kudziwa mulingo woyenera kwambiri ndende?
Kwa IPTG inducer (isopropyl-beta-d-thiogalactoside), kuchulukirachulukira kumakhala bwinoko.Kukhazikika koyenera kumadalira mikhalidwe yeniyeni yoyesera komanso zotsatira zomwe mukufuna.
Nthawi zambiri, kuchuluka kwa IPTG kumagwiritsidwa ntchito mumitundu ya 0.1-1 mm.Kutsika pang'ono kumatha kuchepetsa zotsatira zoyipa pakukula kwa ma cell ndipo kumatha kuchepetsa cytotoxicity chifukwa chakuchulukirachulukira kwa mapuloteni omwe mukufuna.Kuchulukirachulukira kungayambitse kulemedwa kwambiri kwa ma cell, kusokoneza kukula kwa ma cell komanso magwiridwe antchito.
Njira yodziwira kukhazikika koyenera ingakhale kuyesa kuchuluka kwa mapuloteni omwe akuwunikira poyesa kuyesa kwa IPTG mosiyanasiyana.Mayeso ang'onoang'ono azikhalidwe amatha kuyesedwa pogwiritsa ntchito kuchuluka kwa IPTG (monga 0.1 mM, 0.5 mM, 1 mM, ndi zina zambiri.) komanso momwe mawuwo amamvekera pazigawo zosiyanasiyana akhoza kuyesedwa pozindikira kuchuluka kwa mawu a protein yomwe mukufuna (monga Western kuzindikira kwa blot kapena fluorescence).Malingana ndi zotsatira zoyesera, ndende yokhala ndi mawu abwino kwambiri inasankhidwa ngati ndende yabwino kwambiri.
Kuphatikiza apo, mutha kutchulanso zolemba zoyenera kapena zomwe zachitika m'ma laboratories ena kuti mumvetsetse kuchuluka kwa IPTG komwe amagwiritsidwa ntchito nthawi zambiri pansi pamiyeso yofananira, kenako kukhathamiritsa ndikusintha molingana ndi zosowa zoyeserera.
Ndikofunikira kuzindikira kuti kuwongolera koyenera kumatha kusiyanasiyana malinga ndi machitidwe osiyanasiyana ofotokozera, mapuloteni omwe mukufuna, ndi mikhalidwe yoyesera, kotero ndikwabwino kukhathamiritsa pazochitika ndi milandu.
Nthawi yotumiza: Sep-28-2023