I-4-Nitrophenyl beta-D-galactopyranoside CAS:200422-18-0

Umthelela: I-ONPG iyi-substrate esetshenziswa ngokukhethekile ukuthola ubukhona nomsebenzi we-enzyme i-β-galactosidase.Uma i-enzyme ye-β-galactosidase ikhona futhi isebenza, ihlukanisa i-ONPG ibe yimikhiqizo emibili: i-o-nitrophenol kanye nokuphuma kwe-galactose.Ukukhululwa kwe-o-nitrophenol kuphumela ekushintsheni kombala ophuzi, okungalinganiswa kusetshenziswa i-spectrophotometer.

Isicelo: I-ONPG inezinhlelo zokusebenza eziningana ku-molecular biology kanye nocwaningo lwe-biochemistry:

Ukunqunywa komsebenzi we-β-galactosidase: I-ONPG ivamise ukusetshenziselwa ukukala kanye nokulinganisa umsebenzi we-enzyme ye-β-galactosidase.Izinga lokwakheka kwe-o-nitrophenol, elilingana ngokuqondile nomsebenzi we-enzyme, lingalinganiswa nge-spectrophotometrically.

Isisho sofuzo kanye nomthethonqubo: I-ONPG ivame ukusetshenziswa ekuhlolweni okuhlobene nokuvezwa kofuzo kanye nezifundo zokulawula.Ivame ukusetshenziswa ekuhlolweni kwamaprotheni e-fusion, njengesistimu ye-fusion ye-lacZ evame ukusetshenziswa, ukutadisha ukubonakaliswa kwezakhi zofuzo ngaphansi kokulawulwa ngabagqugquzeli abathile.Umsebenzi we-beta-galactosidase okalwa kusetshenziswa i-ONPG unikeza imininingwane ngezinga lenkulumo yofuzo.

Ukuhlolwa komsebenzi we-β-galactosidase: I-ONPG ingasetshenziswa njengendlela yokuhlola i-colorimetric kubuchwepheshe be-DNA obuhlangene ukuze kuhlonzwe ubukhona kanye nokusebenza kofuzo lwe-LacZ, olufaka i-β-galactosidase.Le ndlela yokuhlola isiza ekuboneni ama-clones aqukethe isakhi sentshisekelo.

Izifundo ze-Enzyme kinetics: I-ONPG iyasiza futhi ekutadisheni i-kinetics ye-enzyme ye-β-galactosidase.Ngokulinganisa izinga lokusabela kwe-enzyme-substrate ekugxilweni kwe-substrate ehlukene, kungenzeka ukunquma imingcele ye-kinetic efana ne-Michaelis-Menten constants (Km) kanye namazinga aphezulu okuphendula (Vmax).