包含体表达基因工程牛生长激素提取、变复性及纯化的研究

Study on the extraction,denaturation,renaturation and purification of inclusion body-expressed bovine growth hormone

为了取得较好的复性效果,对重组牛生长激素的纯化及变复性条件进行了研究。结果表明:发酵菌体高压匀浆破碎、离心得到的包含体在7.5 mol/L尿素中变性24 h后,以100 mmol/L Tris.HCl pH10.0稀释至尿素浓度4.5 mol/L,4℃复性48 h,对20 mmol/LTris.HCl pH9.0透析除盐可以成功对重组牛生长激素复性;采用截留分子量100 kD、5 kD超滤膜对复性重组牛生长激素浓缩,FPLC纯化后可得到高纯度的目标蛋白。

The conditions of purification, denaturation and renaturation of recombinant Bovine growth hormone（BGH） were researched in order to get a good renaturation result. The fermentative thalluses were disrupted through high-pressure homogenization, and the inclusion body obtained by centrifugalization was denatured in 7.5 mol/L urea for 24 h, diluted with 100 mmol/L Tris · HCl at pH 10. 0 until the urea concentration was 4. 5 mol/L, and renatured at 4℃ for 48 h. Dialysing 20 mmol/L Tris · HCl at pH 9.0 and removing salt could successfully renature the recombinant BGH. The renatured recombinant BGH was concentrated with 100 K and 5 K ultrafilters and purified by fast protein liguid chromatography（FPLC）, and thus the target protein of high purity could be got.