Regulated-Integration for the Sequential Insertion of Metabolic Pathway and Product Genes in Laboratory and Industrial Yeast Strains

9708299 DaSilva酵母Saccharomyces cerevisae是一种非常重要的工业微生物，也是了解真核细胞过程的关键实验室微生物。 所采用的菌株范围从遗传上充分表征的单倍体或二倍体菌株到相对不表征的工业多倍体菌株。 以最佳拷贝数引入稳定的外源基因的能力是极其重要的，并且设计这种酵母的代谢的能力具有巨大的潜力。 还没有开发出以精确拷贝数稳定插入一系列相同或不同基因的简单方法，并且还没有解决将多个基因调节插入工业上重要的多倍体酵母中的问题。 该项目通过开发和应用实验室和工业（包括多倍体）酵母菌株中途径和产物基因的顺序整合方法来满足这些需求。 所获得的知识将有助于重组酵母表达系统的改进，并可能提供深入了解高等真核细胞的整合方法。 ***

9708299 DaSilva The yeast Saccharomyces cerevisae is a very important industrial microorganism, and a key laboratory microorganism for understanding eucaryotic cell processes. The strains employed range from genetically well-characterized haploid or diploid strains to the relatively uncharacterized industrial polyploid strains. The ability to introduce stable foreign genes at optimum copy number is extremely important, and the ability to engineer the metabolism of this yeast has tremendous potential. Straightforward methods to stably insert a series of the same or different genes at precise copy numbers have not been developed, and the regulated insertion of multiple genes into the industrially important polyploid yeast has not been addressed. This project addresses these needs by developing and applying methods for the sequential integration of pathway and product genes in both laboratory and industrial (including polyploid) yeast strains. The knowledge to be obtained is to contribute to the improvement of recombinant yeast expression systems, and may provide insight into integration methods of higher eucaryotic cells. ***