4-Nitrofenyl beta-D-galactopyranoside CAS:200422-18-0

Saamaynta: ONPG waa substrate si gaar ah loo isticmaalo si loo ogaado joogitaanka iyo dhaqdhaqaaqa enzyme β-galactosidase.Marka β-galactosidase enzyme uu jiro oo uu firfircoon yahay, waxay ONPG u kala jajabisaa laba badeeco: o-nitrophenol iyo derivative galactose.Xoraynta o-nitrophenol waxay keentaa isbeddel midab huruud ah, kaas oo lagu cabbiri karo iyadoo la isticmaalayo spectrophotometer.

Codsiga: ONPG waxay leedahay codsiyo dhowr ah oo ku saabsan bayoolaji molecular iyo cilmi-baarista biochemistry:

Go'aaminta waxqabadka β-galactosidase: ONPG waxaa badanaa loo isticmaalaa in lagu cabbiro laguna qiyaaso waxqabadka β-galactosidase enzyme.Heerka samaynta o-nitrophenol, taas oo si toos ah u dhiganta dhaqdhaqaaqa enzyme, waxaa lagu qiyaasi karaa spectrophotometrically.

Muujinta hidda-wadaha iyo habaynta: ONPG waxaa inta badan loo adeegsadaa tijaabooyinka la xidhiidha muujinta hidda-wadaha iyo daraasaadka sharciga.Waxaa si joogta ah loo isticmaalaa baaritaanka borotiinka fiyuuska, sida nidaamka fiyuuska lacZ ee inta badan la isticmaalo, si loo barto muujinta hiddo-wadaha hoos yimaada xakamaynta dhiirrigeliyeyaasha gaarka ah.Hawsha beta-galactosidase ee lagu cabbiro iyadoo la isticmaalayo ONPG waxay bixisaa aragtiyo ku saabsan heerka muujinta hiddasidaha.

Baaritaanka waxqabadka β-galactosidase: ONPG waxaa loo isticmaali karaa habka baarista midabka midabka ee tignoolajiyada dib-u-dhiska DNA si loo aqoonsado joogitaanka iyo shaqeynta hidda-wadaha LacZ, kaas oo qeexaya β-galactosidase.Habkan baadhistu waxa uu caawinayaa in la aqoonsado clones ka kooban hidda-wadaha xiisaha.

Daraasadaha kinetics enzyme: ONPG sidoo kale waxay faa'iido u leedahay barashada kinetics ee β-galactosidase enzyme.Marka la cabbiro heerka falcelinta enzyme-substrate-ka ee isku-ururinta substrate-ka kala duwan, waxaa suurtagal ah in la go'aamiyo cabbirrada kinetic sida Michaelis-Menten joogto ah (Km) iyo heerka falcelinta ugu badan (Vmax).