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

Isiphumo: I-ONPG sisityana esisetyenziswa ngokukodwa ukubona ubukho kunye nomsebenzi we-enzyme i-β-galactosidase.Xa i-enzyme ye-β-galactosidase ikhona kwaye isebenza, iqhekeza i-ONPG kwiimveliso ezimbini: i-o-nitrophenol kunye ne-galactose derivative.Ukukhululwa kwe-o-nitrophenol kubangela utshintsho lombala ophuzi, onokulinganiswa usebenzisa i-spectrophotometer.

Isicelo: I-ONPG inezicelo ezininzi kwibhayoloji yemolekyuli kunye nophando lwebhayoloji:

Ukuzimisela komsebenzi we-β-galactosidase: I-ONPG isetyenziswa ngokuqhelekileyo ukulinganisa kunye nokulinganisa umsebenzi we-enzyme ye-β-galactosidase.Umlinganiselo wokwenziwa kwe-o-nitrophenol, ehambelana ngokuthe ngqo nomsebenzi we-enzyme, unokulinganiswa nge-spectrophotometrically.

Ukubonakaliswa kofuzo kunye nommiselo: I-ONPG ihlala isetyenziswa kuvavanyo olunxulumene nokubonakaliswa kofuzo kunye nezifundo zolawulo.Isetyenziswa rhoqo kwiimvavanyo zeprotheyini ye-fusion, njengenkqubo esetyenziswa ngokuqhelekileyo ye-lacZ fusion, ukufunda ukubonakaliswa kwemfuza phantsi kolawulo lwabakhuthazi abathile.Umsebenzi we-beta-galactosidase ulinganiswe kusetyenziswa i-ONPG ibonelela ngengqiqo kwinqanaba le-gene expression.

Ukuhlolwa komsebenzi we-β-galactosidase: I-ONPG ingasetyenziselwa njengendlela yokuhlola i-colorimetric kwi-teknoloji ye-DNA edibeneyo ukuchonga ubukho kunye nokusebenza kwe-gene ye-LacZ, efaka i-β-galactosidase.Le ndlela yokuhlola inceda ekuchongeni ii-clones eziqulethe imfuza yomdla.

Izifundo ze-enzyme kinetics: I-ONPG ikwaluncedo ekufundeni i-kinetics ye-enzyme ye-β-galactosidase.Ngokulinganisa umlinganiselo we-enzyme-substrate reactions kwiindawo ezahlukeneyo ze-substrate, kunokwenzeka ukugqiba i-kinetic parameters ezifana ne-Michaelis-Menten constants (Km) kunye nemilinganiselo ephezulu yokuphendula (Vmax).