Expression of human alcohol dehydrogenase ADH1B2 in Pichia pastoris and its activity analysis(人乙醇脱氢酶ADH1B2在毕赤酵母中的表达及活性分析)

TANGTian-tian(汤甜甜); ZHOULiang(周亮); JIPeng-fei(吉鹏飞); BAOWen-na(鲍文娜); GONGXing-guo(龚兴国)

doi:10.3785/j.issn.1008-9497.2012.05.013

Zhejiang Daxue xuebao. Lixue ban (Sep 2012)

Expression of human alcohol dehydrogenase ADH1B2 in Pichia pastoris and its activity analysis(人乙醇脱氢酶ADH1B2在毕赤酵母中的表达及活性分析)

TANGTian-tian(汤甜甜),
ZHOULiang(周亮),
JIPeng-fei(吉鹏飞),
BAOWen-na(鲍文娜),
GONGXing-guo(龚兴国)

Affiliations

TANGTian-tian(汤甜甜): Institute of Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China(浙江大学生命科学学院，生物化学研究所，浙江杭州 310058)
ZHOULiang(周亮): Institute of Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China(浙江大学生命科学学院，生物化学研究所，浙江杭州 310058)
JIPeng-fei(吉鹏飞): Institute of Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China(浙江大学生命科学学院，生物化学研究所，浙江杭州 310058)
BAOWen-na(鲍文娜): Institute of Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China(浙江大学生命科学学院，生物化学研究所，浙江杭州 310058)
GONGXing-guo(龚兴国): Institute of Biochemistry, College of Life Science, Zhejiang University, Hangzhou 310058, China(浙江大学生命科学学院，生物化学研究所，浙江杭州 310058)

DOI: https://doi.org/10.3785/j.issn.1008-9497.2012.05.013
Journal volume & issue: Vol. 39, no. 5
pp. 557 – 563

Abstract

Read online

根据GeneBank中人乙醇脱氢酶ADH1B2基因序列设计引物，以人肝脏总RNA为模板，反转录获得人乙醇脱氢酶基因.将克隆基因与表达载体pPICZB连接，测序正确的pPICZB-ADH1B2电转化毕赤酵母GS115；重组毕赤酵母工程菌经0.5%(v/v)甲醇诱导72 h后目的蛋白以可溶形式表达，经DEAE-Sepharose FF纯化后蛋白酶活性可达8 000 U • mg-1.同时构建重组质粒pCDNA3.1-ADH1B2转染HepG2人肝癌细胞，发现细胞内ROS/GSH水平降低，Akt磷酸化水平降低，细胞活性下降.为临床酒精中毒预防和治疗药物的研发以及ADH在肝脏肿瘤方面的研究提供了理论基础.

Published in Zhejiang Daxue xuebao. Lixue ban

ISSN: 1008-9497 (Print)
Publisher: Zhejiang University Press
Country of publisher: China
LCC subjects: Science: Mathematics: Instruments and machines: Electronic computers. Computer science; Science: Physics
Website: https://www.zjujournals.com/sci/EN/1008-9497/home.shtml

About the journal

Abstract

Keywords