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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2012, Vol. 38 Issue (1): 43-47    DOI: 10.3785/j.issn.1008-9209.2012.01.006
生物科学与技术     
基于α半乳糖苷酶基因为筛选标记的酿酒酵母诱导型表达载体的构建及应用
张伟１ , 郭钦２ , 阮辉３ , 何国庆３
１ .温州医学院基础医学院,浙江温州３２５０３５ ; 2.江苏大学食品学院,江苏镇江２１２０１３ ;
３ .浙江大学生物系统工程与食品科学学院,浙江杭州３１００５８
Construction of an inducible expression vector based on αgalactosidase gene as a selection marker forSaccharomyces cerevisiae and its application
ZHANG Wei１ , GUO Qin２ , RUAN Hui３ , HE Guo‐qing３
1 .School o f Basic Medical Science , Wenzhou Medical College , Wenz hou , Zhej iang 325035 , China ; 2 .School o f Food and Biological Engineering ,
Jiangsu University , Zhenj iang , Jiangsu 212013 , China ; 3 .School o f Biosystems Engineering and Food Science , Zhejiang University , H angz hou 310058 , China
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摘要： 以α‐半乳糖苷酶基因为筛选标记,构建GAL１ 诱导型启动子介导的酿酒酵母表达载体YGM‐α‐gal 质粒,将枯草芽孢杆菌的β‐１ ,３‐１ ,４‐葡聚糖酶基因克隆到此载体中,构建质粒YGMPA‐α‐gal ,转化宿主酵母后,实现β‐１ ,３‐１ ,４‐葡聚糖酶在酿酒酵母中的分泌表达.结果表明:在２％ 半乳糖诱导下,摇瓶发
酵２４ h 后分泌表达的β‐葡聚糖酶活性达到４１１.９ U.mL － １ ,而在培养６０ h 后,发酵液中α‐半乳糖苷酶活性可达６４.２ U.mL － １ .说明α‐半乳糖苷酶基因可用作酿酒酵母表达载体转化的有效筛选标记,为食品级酿酒酵母表达系统的构建提供了新选择.
Abstract: YGM‐α‐gal , an expression vector for Saccharomyces cerev isiae , was constructed containing α‐
galactosidase gene as selection marker and an inducible GAL１ promoter . The β‐１ ,３‐１ ,４‐glucanase gene
cloned from Bacillus subtilis was inserted into the multiple cloning sites of the plasmid YGM‐α‐gal to
generate YGMPA‐α‐gal , then to transform the host yeast , and β‐１ ,３‐１ ,４‐glucanase was efficiently
expressed as a secretive protein in S . cerev isiae . The results indicated that the β‐１ ,３‐１ ,４‐glucanase
activity in the ferment liquid in shake flask was about ４１１.９ U.mL -１ after adding ２％ galactose for ２４ h ,
and the α‐galactosidase activity reached ６４.２ U.mL -１ after adding ２％ galactose for ６０ h . So the α‐
galactosidase gene can be used as an effective selection marker for screening the positive transformants of
food‐grade yeast expression system .
出版日期: 2012-01-20
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张伟
郭钦
阮辉
何国庆

引用本文:

张伟,郭钦,阮辉,何国庆. 基于α半乳糖苷酶基因为筛选标记的酿酒酵母诱导型表达载体的构建及应用[J]. 浙江大学学报(农业与生命科学版), 2012, 38(1): 43-47.

ZHANG Wei,GUO Qin,RUAN Hui,HE Guo qing. Construction of an inducible expression vector based on αgalactosidase gene as a selection marker forSaccharomyces cerevisiae and its application. , 2012, 38(1): 43-47.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2012.01.006    或    http://www.zjujournals.com/agr/CN/Y2012/V38/I1/43

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