谷氨酰胺转胺酶在毕赤酵母菌的初步表达  

The Preliminary Expression of Transglutaminase Gene in pichia pastoris

作　　者：何冬兰[1] 叶程[1] 王亚南[1] 覃桂[1] 

机构地区：[1]中南民族大学生命科学学院,武汉430074

出　　处：《中南民族大学学报（自然科学版）》2013年第1期32-35,共4页Journal of South-Central University for Nationalities：Natural Science Edition

基　　金：湖北省自然科学基金资助项目(2010CDZ046)

摘　　要：从重组菌株pPIC3.5k-MTG/DH5α提取重组质粒pPIC3.5k-MTG,经Sac I酶切线性化处理后纯化重组质粒电击转化到毕赤酵母菌中,用甲醇诱导表达,经SDS-PAGE鉴定重组质粒P.pastoris GS115成功表达出了TGase蛋白,并初步纯化获得了较纯的目的蛋白,其表观分子量为47 kD,酶活为0.50 U/mL.Reconstructed expression plasmid pPIC3.5k-MTG was obtained from recombinant strain pPIC3.5k-MTG/DH5α.After lineared by Sac I digesting,plasmid was transformed into pichia pastoris GS115 through electroporation.The recombinant P.pastoris GS115 was inducted by methanol.SDS-PAGE analysis showed that the recombinant P.pastoris GS115 expressed tracelular protein with apparent molecular weight of 47 kD and enzyme activity of 0.50 U/mL by preliminary purification.

关 键 词：谷氨酰胺转胺酶 毕赤酵母 表达 

分 类 号：Q556[生物学—生物化学]