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

Cov nyhuv: ONPG yog ib qho substrate tshwj xeeb uas siv los txheeb xyuas qhov muaj thiab kev ua haujlwm ntawm enzyme β-galactosidase.Thaum β-galactosidase enzyme yog tam sim no thiab nquag, nws cleaves ONPG rau hauv ob yam khoom: o-nitrophenol thiab galactose derivative.Kev ywj pheej ntawm o-nitrophenol ua rau muaj xim daj hloov, uas tuaj yeem ntsuas tau siv lub spectrophotometer.

Daim Ntawv Thov: ONPG muaj ntau daim ntawv thov hauv molecular biology thiab biochemistry tshawb fawb:

Kev txiav txim siab ntawm β-galactosidase kev ua haujlwm: ONPG feem ntau siv los ntsuas thiab ntsuas qhov kev ua ntawm β-galactosidase enzyme.Tus nqi ntawm o-nitrophenol tsim, uas yog ncaj qha proportional rau cov enzyme kev ua, yuav ntsuas spectrophotometrically.

Gene qhia thiab kev tswj hwm: ONPG feem ntau siv rau hauv kev sim cuam tshuam nrog cov noob qhia thiab kev tshawb fawb txog kev cai.Nws tau nquag siv hauv kev tshuaj ntsuam fusion protein, xws li kev siv lacZ fusion system, los kawm txog kev qhia ntawm cov noob nyob rau hauv kev tswj hwm ntawm cov neeg txhawb nqa tshwj xeeb.Kev ua haujlwm beta-galactosidase ntsuas siv ONPG muab kev nkag siab rau qib ntawm cov noob qhia.

Kev tshuaj ntsuam rau β-galactosidase kev ua ub no: ONPG tuaj yeem siv los ua kev tshuaj ntsuam xim xim hauv DNA thev naus laus zis los txheeb xyuas qhov muaj thiab ua haujlwm ntawm LacZ gene, uas encodes β-galactosidase.Txoj kev tshuaj ntsuam no pab txhawm rau txheeb xyuas cov clones uas muaj cov noob ntawm kev txaus siab.

Enzyme kinetics kev tshawb fawb: ONPG kuj tseem muaj txiaj ntsig hauv kev kawm cov kinetics ntawm β-galactosidase enzyme.Los ntawm kev ntsuas tus nqi ntawm cov enzyme-substrate cov tshuaj tiv thaiv ntawm qhov sib txawv substrate concentrations, nws muaj peev xwm txiav txim siab kinetic tsis xws li Michaelis-Menten tas mus li (Km) thiab qhov siab tshaj plaws cov tshuaj tiv thaiv (Vmax).