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

Umphumela: I-ONPG iyi-substrate esetshenziselwa ngqo ukuthola ubukhona kanye nomsebenzi we-enzyme i-β-galactosidase. Uma i-enzyme ye-β-galactosidase ikhona futhi isebenza, ihlukanisa i-ONPG ibe imikhiqizo emibili: i-o-nitrophenol kanye ne-galactose derivative. Ukukhululwa kwe-o-nitrophenol kuphumela ekushintsheni kombala ophuzi, okungalinganiswa kusetshenziswa i-spectrophotometer.

Isicelo: I-ONPG inezinhlelo zokusebenza eziningana ocwaningweni lwe-molecular biology kanye ne-biochemistry:

Ukunqunywa komsebenzi we-β-galactosidase: I-ONPG ivame ukusetshenziswa ukukala nokulinganisa umsebenzi we-enzyme ye-β-galactosidase. Izinga lokwakheka kwe-o-nitrophenol, elilingana ngqo nomsebenzi we-enzyme, lingalinganiswa nge-spectrophotometric.

Ukuvezwa kwezakhi zofuzo kanye nokulawulwa kwazo: I-ONPG ivame ukusetshenziswa ezivivinyweni ezihlobene nokuvezwa kwezakhi zofuzo kanye nezifundo zokulawulwa kwazo. Ivame ukusetshenziswa ekuhlolweni kwamaprotheni okuhlanganiswa, njengohlelo lokuhlanganiswa kwe-lacZ olusetshenziswa kakhulu, ukutadisha ukuvezwa kwezakhi zofuzo ngaphansi kokulawulwa kwabagqugquzeli abathile. Umsebenzi we-beta-galactosidase olinganiswa kusetshenziswa i-ONPG unikeza ukuqonda ngezinga lokubonakaliswa kwezakhi zofuzo.

Ukuhlolwa komsebenzi we-β-galactosidase: I-ONPG ingasetshenziswa njengendlela yokuhlola i-colorimetric kubuchwepheshe be-DNA ehlanganisiwe ukuhlonza ukuba khona kanye nokusebenza kwe-gene ye-LacZ, efaka i-β-galactosidase. Le ndlela yokuhlola isiza ekuboneni ama-clone aqukethe i-gene ethakaselwayo.

Izifundo ze-kinetics ze-enzyme: I-ONPG nayo iwusizo ekutadisheni i-kinetics ye-enzyme ye-β-galactosidase. Ngokulinganisa izinga lokusabela kwe-enzyme-substrate ekugxilweni okuhlukene kwe-substrate, kungenzeka ukunquma amapharamitha e-kinetic njenge-Michaelis-Menten constants (Km) kanye ne-maximum reaction rates (Vmax).