利用油体蛋白体外重组技术实现牛肠激酶固定化应用  

Immobilized Application of Bovine Erokinae by in Vitro Oleosin Recombination Technology

作　　者：董立强[1,2] 李梅[2] 张燕燕[1,2] 孙文丽[2] 陈红漫[1] 刘昱辉[2] 

机构地区：[1]沈阳农业大学生物科学技术学院,沈阳110161 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《湖北农业科学》2011年第4期849-853,共5页Hubei Agricultural Sciences

基　　金：国家"863"计划项目(2007AA100503)

摘　　要：为研究利用油体蛋白体外重组技术实现肠激酶固定化应用的可行性,利用油体蛋白Oleosin、自剪切蛋白Intein和人工合成的牛肠激酶轻链(Enterokinase,EK)cDNA,分别构建表达盒Intein-oleosin-EK和Intein-EK,插入毕赤酵母表达载体pPIC9K,转化毕赤酵母GS115,甲醇诱导表达后通过SDS-PAGE和Western-blot检测发现,带有Intein-肠激酶表达盒的阳性菌株中有目的蛋白的表达,而Oleosin-intein-肠激酶融合蛋白表达盒不能在酵母中表达。To study the possibility of immobilized application of enterokinae（EK） by artificial oil bodies,two expression cassettes,oleosin-intein-EK and intein-EK,consisting of oleosin gene,intein and artificially synthesized EK cDNA were constructed.The expression cassettes were then inserted into pPIC9K and transformed into Pichia pastoris GS115 strain.After induced by methanol,SDS-PAGE and Western blot were used to test the expression products;and analysis showed that target protein was expressed in the strains containing intein-EK expression cassette,while oleosin-intein-EK was not expressed.

关 键 词：油体蛋白 人工油体 肠激酶 固定化酶 

分 类 号：Q556.9[生物学—生物化学] Q814.2