Flagfeldt Dongmei Bai, Siewers Verena, Huang Le, Nielsen Jens
Centre for Microbial Biotechnology, Department of Systems Biology, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.
Yeast. 2009 Oct;26(10):545-51. doi: 10.1002/yea.1705.
The construction of mitotically stable yeast strains for heterologous gene or pathway expression often requires chromosomal integration. However, transcription levels vary between different chromosome regions. We therefore characterized 20 different integration sites of the Sacchromyces cerevisiae genome by inserting lacZ as a reporter gene under the control of two different promoters and determining expression levels through enzyme activity measurement. An up to 8.7-fold difference was detected between the sites conferring lowest and highest expression, respectively. This opens the opportunity for modulating gene expression levels while retaining promoter and culture conditions.
构建用于异源基因或途径表达的有丝分裂稳定酵母菌株通常需要进行染色体整合。然而,不同染色体区域之间的转录水平存在差异。因此,我们通过在两种不同启动子的控制下插入lacZ作为报告基因,并通过酶活性测量来确定表达水平,从而对酿酒酵母基因组的20个不同整合位点进行了表征。在分别赋予最低和最高表达的位点之间检测到高达8.7倍的差异。这为在保留启动子和培养条件的同时调节基因表达水平提供了机会。