鼠肝中金属硫蛋白的纯化.pdf

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 3 王雪 赵海峰 吕明 李鸿雁 吴文杰 刘娅 李正强 (, 130021) : 旨在探索鼠肝中金属硫蛋白(MT )提取 艺并加以改进按照经典方法 艺提取 MT,用中空纤维柱超滤方法 加以改进, 依据MT 自身特点进行分离和鉴定结果显示, 粗品符合 MT特点: 分子量在 65 kD 左右; 无 280 nm特征吸收峰, 加酸后紫外吸收 (200- 300 nm)肩峰消失; 通过离子交换可以实现 MT 1MT2的分离; 通过原子吸收测定, 含有锌铜镉 3种 金属, 锌含量最高, 具有重要的病理生理意义因此, 与其它方法比较, 改进后方法更适宜实验室制备 : 金属硫蛋白 大鼠 肝脏 层析 The Purification Technology M etallothionein in R at L iver W ang Xue ZhaoHaifeng vM ing iHongyan W uW enjie iu Ya iZhengqiang (J il in Unive rsity, Chang chun 130021) A bstrac:t Itwas to purify them etalloth ionein from the livers ofm ature rats by typ icalprocedures andm ake som e mi provem ents. MT w as isolated by using trad itionalm ethodwh ichw as mi proved hollow- fiber ultrafiltration. Results show ed that the extraction of pro tein molecu lar w eightw as around 65 kD, w ithout 280 nm peak and the absorption shou lder d isappeared( 200- 300 nm)w hen acid add ed. The crude protein could be separated the two form s through DEAE chrom atography. The atom ic absorption analysis show ed that the protein contained copper, cadm ium, and zinc the am ounts ofwhichw as highest. Thus, it proved that the mi provedm ethod is su itable for laboratory preparation. K ey w ords: M etalloth ionein Rat iver Chrom atography (metallothionein, MT) 1 (AgAuBiCdHgZn) 11 供试材料 ( 6- 7 kD), MT W istarTris( [ 1] , ), R ivanol( ), (), DE MT AE, Sephadex G75 ( Amersham Pharmacia Biotech AB)JJ2( , , ), J25( Beckm an Coul MT