Laoteng Kobkul, Cheevadhanarak Supapon, Tanticharoen Morakot, Maresca Bruno
Biochemical Engineering and Pilot Plant Research and Development Unit, National Center for Genetic Engineering and Biotechnology (BIOTEC), King Mongkut's University of Technology Thonburi, Bangkhuntien, Bangkok 10150, Thailand.
Biochem Biophys Res Commun. 2005 Sep 23;335(2):400-5. doi: 10.1016/j.bbrc.2005.07.094.
Promoter study was performed to understand the transcriptional control of delta9-desaturase gene of Mucor rouxii. Several putative cis-elements involved in lipid metabolism were mapped by computational analysis. 5' deletion analysis shows the presence of elements with repressing activity, especially in 122 bp located upstream of the transcription start site. Truncation of these repressor domains showed that the promoter of M. rouxii is functional in Saccharomyces cerevisiae without additional components and is insensitive to nutritional depletion. The promoter also drove effectively the expression of a M. rouxii delta12-desaturase gene, and the linoleic acid content increased with the age of the yeast culture in parallel with the promoter activity. This approach provides a genetic tool for programming heterologous protein production in the yeast.
开展启动子研究以了解鲁氏毛霉Δ9-去饱和酶基因的转录调控。通过计算分析确定了几个参与脂质代谢的假定顺式作用元件。5'缺失分析表明存在具有抑制活性的元件，尤其是在转录起始位点上游122 bp处。这些阻遏结构域的截短表明，鲁氏毛霉的启动子在没有其他组分的情况下在酿酒酵母中具有功能，并且对营养物质耗尽不敏感。该启动子还有效地驱动了鲁氏毛霉Δ12-去饱和酶基因的表达，并且亚油酸含量随着酵母培养时间的延长而增加，与启动子活性平行。这种方法为在酵母中编程异源蛋白生产提供了一种遗传工具。
