Fluorescence resonance zog hloov pauv (FRET)
Fluorescence resonance zog hloov (FRET) yog ib qho uas tsis yog-radiative zog hloov cov txheej txheem nyob rau hauv uas tus neeg mob siab lub xeev lub zog yog pauv mus rau lub receiver zoo siab lub xeev los ntawm kev sib cuam tshuam ntawm intermolecular hluav taws xob ob peb.Cov txheej txheem no tsis cuam tshuam nrog photons thiab yog li tsis muaj hluav taws xob.Qhov kev ntsuam xyuas no muaj qhov zoo ntawm kev ceev, rhiab thiab yooj yim.
Cov xim uas siv hauv FRET qhov kev ntsuam xyuas tuaj yeem zoo ib yam.Tab sis nyob rau hauv feem ntau daim ntawv thov, sib txawv dyes yeej siv.Nyob rau hauv luv luv, kev hloov ntawm luminous resonance zog yog kev hloov ntawm ib khub ntawm dipoles los ntawm tus neeg pub dawb (dye 1) mus rau tus txais (dye 2) thaum pab pawg neeg pub dawb zoo siab.Feem ntau, cov emission spectrum ntawm pab pawg neeg pub dawb fluorophore sib tshooj nrog qhov nqus spectrum ntawm pab pawg txais."Thaum qhov kev ncua deb ntawm ob lub fluorophores yog qhov tsim nyog (10 - 100 A), kev hloov ntawm fluorophore zog los ntawm tus pub rau tus neeg txais yuav raug soj ntsuam."Cov txheej txheem ntawm lub zog hloov pauv nyob ntawm cov qauv tshuaj ntawm cov receptor:
1. Yog hloov dua siab tshiab rau hauv molecular vibration, uas yog, lub teeb luminous ntawm lub zog hloov pauv ploj mus.(Lub receptor yog lub teeb quencher)
2. Lub emission yog khaus ntau dua li receptor nws tus kheej, ua rau redshift nyob rau hauv lub thib ob fluorescence spectrum. "(Receptors yog luminous emitters).
Cov pab pawg neeg pub dawb (EDANS) thiab cov neeg txais cov noob (DABCYL) sib txuas nrog lub ntuj substrate ntawm HIV protease, thiab thaum lub substrate tsis raug rho tawm, DBCYL tuaj yeem quenle EDANS thiab tom qab ntawd ua tsis tau fluorine.Raws li HIV-1 protease disconnection, EDANS tsis quenched los ntawm DABYL, thiab EDANS luciferases tom qab kuaj tau.Qhov muaj cov tshuaj tiv thaiv protease tuaj yeem raug saib xyuas los ntawm kev hloov pauv hauv fluorescence siv EDANS.
FRET peptides yog cov cuab yeej yooj yim los kawm peptidase nonspecificity.Txij li thaum nws cov txheej txheem tshuaj tiv thaiv tuaj yeem soj ntsuam tas li, nws muab txoj hauv kev yooj yim rau kev kuaj xyuas cov haujlwm enzyme.Lub sheen tsim tom qab hydrolysis ntawm peptide daim ntawv cog lus los ntawm tus pub / txais muab ib qho kev ntsuas ntawm enzyme kev ua ntawm nanomolar concentrations.Thaum FRET peptide tsis zoo, nws qhia tau tias ploj mus sai sai ntawm lub flash sab hauv, tab sis thaum twg peptide daim ntawv cog lus tawm tsam tus neeg pub dawb / tus neeg txais khoom tawg, nws tso tawm lub flash, uas tuaj yeem tshawb pom tsis tu ncua thiab kev ua haujlwm enzyme tuaj yeem ua kom muaj nuj nqis.
Post lub sij hawm: Aug-14-2023