Rekombinanta hemokīnu receptora CCR2 sintēze zāļu vielu. skrīningam

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Rekombinanta hemokīnu receptora CCR2 sintēze zāļu vielu. skrīningam

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Darba mērķis bija panākt rekombinanta šķīstoša cilvēka CCR2 hemokīnu receptora analoga ekspresiju baktēriju šūnās.Rekombinantā šķīstošā CCR2 receptora (rsCCR2) kodējošais DNS fragments tika sintezēts mākslīgi. Konstrukcijā ietilpstošā aminoterminālā domēna, ekstracelulāro cilpu un tām blakusesošo transmembranālo domēnu fragmentu kodējošās sekvences tika savienotas, izmantojot mākslīgus DNS linkerus. rsCCR2 sintēzi baktēriju šūnās BL21 (DE3) izdevās panākt, izmantojot ekspresijas vektoru pCold1. Iegūtais nešķīstošais proteīns tika attīrīts ar metālafīno hromatogrāfiju un refoldēts ar iznākumu 1 mg/ml. Lai apstiprinātu iegūtā rekombinantā proteīna funkcionalitāti, tika izmantots imunoblots un ELISA. Imunoblota rezultāti liecina, ka komerciālas anti-CCR2 antivielas atpazīst rekombinanto proteīnu. Sekmīgai Sandwich ELISA sistēmas izveidei nepieciešama papildus optimizācija, lai panāktu specifisku rsCCR2 saistīšanos un kvantificēšanu.

year	journal	country	edition	language
2013-01-01

https://dspace.lu.lv/dspace/handle/7/22956

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