Characterization of chromosomal integration sites for heterologous gene expression in Saccharomyces cerevisiae
Characterization of chromosomal integration sites for heterologous gene expression in Saccharomyces cerevisiae
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DOI：
10.1002/yea.1705
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发表时间：
2009-10-01
期刊：
影响因子：
2.6
通讯作者：
Nielsen, Jens
中科院分区：
文献类型：
--
作者：
Flagfeldt, Dongmei Bai;Siewers, Verena;Nielsen, Jens
构建有丝分裂稳定的外源基因或途径表达酵母菌株通常需要染色体整合。然而，不同染色体区域的转录水平不同。因此，我们在两个不同的启动子控制下插入lacZ作为报告基因，并通过酶活性测定来确定表达水平，从而表征了酿酒Sacchromyces cerevisiae基因组的20个不同整合位点。在最低和最高表达位点之间分别检测到高达8.7倍的差异。这打开了在保留启动子和培养条件的同时调节基因表达水平的机会。版权所有John Wiley & Sons， Ltd. 2009
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. Copyright (C) 2009 John Wiley & Sons, Ltd.