Department of Molecular Microbiology, Washington University School of Medicine, 660 S. Euclid Avenue, Saint Louis, MO 63110, USA.
Fungal Genet Biol. 2010 Mar;47(3):191-8. doi: 10.1016/j.fgb.2009.11.007. Epub 2009 Nov 26.
Cryptococcus neoformans is an opportunistic fungal pathogen that primarily affects immunocompromised individuals. Reverse genetics is commonly used to identify and characterize genes involved in a variety of cellular processes. In C. neoformans there is a limited set of positive selectable markers available to make gene deletions or other genetic manipulations. This has hampered the application of reverse genetics in this organism. We have adapted the Bacteriophage P1 Cre-loxP system for use in C. neoformans and successfully excised and reused the same drug marker, G418, to make two sequential gene deletions, lac1Delta and cap59Delta, in the same strain. This tool will allow investigators to make multiple sequential gene deletions in the same strain, which should facilitate the analysis of multigene families.
新型隐球菌是一种机会性真菌病原体，主要影响免疫功能低下的个体。反向遗传学常用于鉴定和描述参与各种细胞过程的基因。在新型隐球菌中，可用于基因缺失或其他遗传操作的正向选择标记有限。这阻碍了反向遗传学在该生物中的应用。我们已经将噬菌体 P1 Cre-loxP 系统改编用于新型隐球菌，并成功地切除和重复使用相同的药物标记物 G418，在同一菌株中进行了两次连续的基因缺失，lac1Delta 和 cap59Delta。该工具将允许研究人员在同一菌株中进行多次连续的基因缺失，这应该有助于多基因家族的分析。
