Gurvitz A, Coe J G, Dawes I W
School of Biochemistry and Molecular Genetics, University of New South Wales, Kensington, Australia.
Curr Genet. 1993 Nov;24(5):451-4. doi: 10.1007/BF00351856.
Reporter genes consisting of sporulation-specific promoters fused to lacZ were used as markers to monitor the sporulation pathway of the yeast Saccharomyces cerevisiae. Strains transformed with these lacZ gene fusions expressed beta-galactosidase (assayable on plates using the substrate 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside, X-gal) in a sporulation-dependent manner. Mutagenesis experiments performed on transformed strains resulted in the recovery of a number of novel sporulation mutants. Three classes of mutants were obtained: those which overexpressed the reporter gene under sporulation conditions, those which did not express the gene under any conditions, and those which expressed the gene in vegetative cells not undergoing sporulation. On the basis of the blue colony-colour produced in the presence of X-gal these have been described as superblue, white, and blue vegetative mutants, respectively. These were further characterised using earlier reporter genes and other marker systems. This study established that the multicopy reporter plasmids chosen do not interfere with sporulation; they are valid tools for monitoring the pathway and they provide a way to isolate mutations not readily selected by other markers.
由与lacZ融合的孢子形成特异性启动子组成的报告基因被用作标记，以监测酿酒酵母的孢子形成途径。用这些lacZ基因融合体转化的菌株以孢子形成依赖的方式表达β-半乳糖苷酶（使用底物5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷，X-gal在平板上可检测）。对转化菌株进行的诱变实验导致获得了许多新的孢子形成突变体。获得了三类突变体：在孢子形成条件下过表达报告基因的突变体、在任何条件下都不表达该基因的突变体以及在未进行孢子形成的营养细胞中表达该基因的突变体。根据在X-gal存在下产生的蓝色菌落颜色，这些分别被描述为超蓝、白色和蓝色营养突变体。使用早期的报告基因和其他标记系统对它们进行了进一步表征。这项研究表明，所选择的多拷贝报告质粒不会干扰孢子形成；它们是监测该途径的有效工具，并且提供了一种分离不易被其他标记选择的突变的方法。
