本文選題：類彈性蛋白 + 純化標簽�。� 參考：《生物技術(shù)》2013年05期

【摘要】：目的:研究不同濃度的PEG8000對融合蛋白ELP30-Trx相變溫度的影響。方法:將硫氧還蛋白(Thioredoxin,Trx)基因克隆到pET28-ELP30-Trx表達載體中,經(jīng)IPTG誘導在宿主菌E.coli BLR(DE3)中表達ELP30-Trx,通過可逆相變循環(huán)(Inverse transition cycling,ITC)純化融合蛋白。測量相變溫度(T t),并檢測不同濃度的PEG8000對ELP30-Trx T t的影響。結(jié)果:經(jīng)酶切及測序鑒定證實成功構(gòu)建了ELP30-Trx表達載體,經(jīng)Westen-Blot檢測表明成功表達ELP30-Trx融合蛋白。經(jīng)ITC純化后的ELP30-Trx純度可達95%以上。5%、10%、15%、20%的PEG8000(W/V)使終濃度為25μmol/L ELP30-Trx的T t從37.9℃分別降低至34.6℃、28.5℃、14.0℃、6.0℃。結(jié)論:成功構(gòu)建、表達融合蛋白ELP30-Trx,PEG8000能顯著降低ELP30-Trx相變溫度。通過對類彈性蛋白的研究,為類彈性蛋白的擴大應(yīng)用提供理論依據(jù)。
[Abstract]:Aim: to study the effect of different concentrations of PEG8000 on phase transition temperature of ELP30-Trx fusion protein. Methods: thioredoxin Trx) gene was cloned into pET28-ELP30-Trx expression vector. ELP30-Trx was expressed in host strain E.coli BLRDE3 by IPTG. The fusion protein was purified by reversible phase change cycle reverse transition cycling. The phase transition temperature was measured and the effect of different concentration of PEG8000 on ELP30-Trx T t was measured. Results: ELP30-Trx expression vector was successfully constructed by restriction endonuclease digestion and sequencing, and ELP30-Trx fusion protein was successfully expressed by Westen-Blot. The purity of ELP30-Trx purified by ITC can reach more than 95% and 20% PEG8000W / V). The final concentration of T t of 25 渭 mol/L ELP30-Trx is reduced from 37.9 鈩，

本文編號：1831722