Latour system精简酿酒酵母染色体方法的建立及应用

酿酒酵母是第一个完成全基因组测序的真核生物，具有广泛的科研应用价值。利用酿酒酵母的全基因组序列可以进行精确的基因定位及敲除，从而达到对其基因组进行精简的目的，为合成生物学最小基因组的研究工作打下基础。根据Latour system 设计敲除所需引物，构建敲除盒，筛选重组体和缺失体，成功敲除酿酒酵母a型单倍体染色体XIII中339301-352281 nt包含的8个基因，为酿酒酵母染色体精简奠定基础，同时证明了Latour system 可以应用于酿酒酵母大片段敲除。

Establishment & Application of Chromosome Simplify by Latour System in S. cerevisiae

Saccharomyces cerevisiae is the first eucaryote to complete its whole genome sequencing; it has extensive application value of scientific research. Adopting the whole genome sequencing of S. cerevisiae could precisely conduct its gene location and deletion, accordingly to reach the goal of its genome simplification, therefore, lay down a foundation for the study on synthesis of an organism with minimal genome. In this paper, according to Latour system, primers needed were precisely designed and deleted to construct a deletion cassette, and screen recombinant and deletant. Eight genes containing in 339301 352281 nt in chromosome XIII of monoploid S. cerevisiae type a were successfully deleted, and thereby laid down a foundation for chromosome simplification in S. cerevisiae. At the same time it had proved that Latour system can be applied in large fragment deletion in S. cerevisiae.