Tiknoolajiyada FRET peptide

Wareejinta tamarta resonance fluorescence (FRET)

Wareejinta tamarta resonance fluorescence (FRET) waa habka wareejinta tamarta shucaaca ah kaas oo deeq-bixiyuhu ku faraxsan yahay tamarta gobolka loo wareejinayo aqbalaha gobolka xiisaha leh iyada oo loo marayo isdhexgalka lammaanaha korantada intermolecular.Habkani kuma lug laha sawir-qaadayaasha sidaas darteedna maaha shucaac.Qiimayntani waxay leedahay faa'iidooyinka degdegga, xasaasiga ah iyo fududaanta.

Rinjiga lagu isticmaalo baaritaanka FRET wuxuu noqon karaa isku mid.Laakiin inta badan codsiyada, midabyo kala duwan ayaa dhab ahaantii loo isticmaalaa.Marka la soo koobo, wareejinta tamarta resonance ee iftiinka leh waa wareejinta lammaane dipoles oo ka yimid deeq bixiyaha (dheee 1) aqbalaha (dheeyowga 2) marka kooxda ku-deeqayaashu ay faraxsan yihiin.Guud ahaan, qiiqa qiiqa ee kooxda fluorophore ee Donor waxay ku dul wareegaysaa spectrum nuugista kooxda Aqbalaha."Marka masaafada u dhaxaysa labada fluorophores ay ku habboon tahay (10 - 100 A), wareejinta tamarta fluorophore ee deeq bixiyaha waa la arki karaa."Habka wareejinta tamarta waxay ku xiran tahay qaab dhismeedka kiimikada ee soo-dhoweeyaha:

1. Waxaa loo beddelaa gariir molecular, taas oo ah, iftiinka iftiinka ee wareejinta tamarta wuu baaba'aa.(Qaybiyuhu waa bakhtiiye iftiin leh)

2. Sii dayntu waa ka xoog badan tahay reseptor-ka laftiisa, taas oo keenta isbeddel guduudan oo ku yimaadda spectrum fluorescence secondary.(Receptors waa qiiqa nuurka leh).

Kooxda ku-deeqaha (EDANS) iyo hidda-wadaha aqbalaadda (DABCYL) waxay si isku mid ah ugu xidhan yihiin substrate-ka dabiiciga ah ee borotiinka HIV, iyo marka substrate-ka aan la goyn, DABCYL waxay demi kartaa EDANS ka dibna waxay noqon kartaa mid aan la ogaan karin fluorine.Marka HIV-1 ka go'o borotiinka, EDANS ma damiso DABCYL, ka dibna EDNS luciferases ayaa la ogaan karaa.Helitaanka ka-hortagga borotiinka waxaa lagula socon karaa isbeddelada xoojinta iftiinka iftiinka ee EDNS.

FRET peptides waa qalab ku habboon in lagu barto peptidase aan gaar ahayn.Maadaama habka falcelinta si joogto ah loola socon karo, waxay bixisaa hab ku habboon oo lagu ogaanayo dhaqdhaqaaqa enzyme.Sheen soo saaray ka dib hydrolysis of peptide bonds by ku-deeqaha/aqbalaha ayaa bixisa cabirka dhaqdhaqaaqa ensaymes ee uruurinta nanomolar.Marka peptide-ka FRET uu yahay mid sugan, waxa ay muujinaysaa luminta degdega ah ee fiilada gudaha, laakiin marka xidhid kasta oo peptide ah oo ka soo horjeeda ku-deeqaha/aqbalaha jabo, waxa ay sii daysaa fallaadh, kaas oo si joogto ah loo ogaan karo iyo dhaqdhaqaaqa enzyme ayaa markaa la qiyaasi karaa.