重组蛋氨酸裂解酶的表达、复性及其活性的研究.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 4 1 2 3 2 2 2 孔晨虹 夏立亮 徐顺利 于源华 蒋琴 杨佳新 1 2 3 ( , 130022; , 130022; , 130031) : 探索以包涵体形式表达的重组蛋 酸裂解酶的纯化复性方法, 并对其活性进行检测将阴道毛滴虫蛋 酸裂解酶重组表达载体 PET15bmgl1转化大肠杆菌 BL21, 经异丙基D硫代半乳糖苷 ( IPTG)诱导, 并对表达条件 进行优化, 获得大量表达通过对包涵体的纯化及复性研究, 检测重组蛋 酸裂解酶的免疫活性及酶活性W estern blotting结果表明, 重组蛋 酸裂解酶免疫小鼠制备的多抗可以和从阴道毛滴虫中提取的天然蛋 酸裂解酶发生特异性 反应活性检测结果显示复性蛋 酸裂解酶有活性, 复性效率达到 25 左右以包涵体形式表达的蛋 酸裂解酶经 变性纯化及复性后, 获得大量有活性的酶, 为深入了解其结构功能及酶学性质, 开展其在临床检测中的应用研究奠定 基础 : 蛋 酸裂解酶 同型半胱 酸 基因表达 包涵体 复性 Expression, Renaturation and Activity of RecombinantM ethionine Lyase Expressed as Inclusion Body inE. coli 1 2 3 2 2 2 Kong Chenhong XiaLiliang Xu Shunli Yu Yuanhua JiangQin Yang Jiaxin 1 ( Trad itiona l ChineseM ed ic ineH osp ita l of Ch ang chun, Cha ng chun 130022; 2 Colleg e of Lfi e Sc ience, Chang chun Un iv ersity of Ssc ience and T ech nology, Chang chun 130022; 3 E rdao D istrictH osp ital in Chang chun, Chang ch un 130031) Abstrac:t Itwas to study the purification, renaturation and activity of recombinantMethionine Lyase expressed as inclusion body inE coli. TheE coli BL21 transformedw ith PET15bmgl1was cultured and induced by IPTG. The best expression condition of recombinantMethionine Lyasewas decided by screening. The purification and renaturation of recombinantMethionine Lyasewere explored and the activitywastested. Itwas proved byWestern blot