Moreau A, Durand S, Morosoli R
Centre de Recherche en Microbiologie Appliquée, Institut Armand-Frappier, Ville de Laval, Québec, Canada.
Gene. 1992 Jul 1;116(1):109-13. doi: 10.1016/0378-1119(92)90637-5.
The xylanase(XLN)-encoding gene(XLN) of Cryptococcus albidus and its cDNA were each inserted into the vector, pVT100, for expression in Saccharomyces cerevisiae. Expression was under the control of either their own promoter or the gene encoding alcohol dehydrogenase (ADH1) promoter. Yeast transformed with plasmids containing the cDNA of the structural XLN gene and the XLN promoter produced active extracellular XLN when grown with galactose as carbon source. However, with glucose as carbon source, XLN was repressed. Using the ADH1 promoter, which is stimulated by glucose, XLN was secreted into the culture medium. In both cases, the secreted 48-kDa enzyme corresponded to the native XLN produced by C. albidus. With the plasmid bearing the genomic XLN gene, there was transcription, but the seven introns interrupting XLN were not spliced out by S. cerevisiae and no enzyme was produced.
将浅白隐球酵母的木聚糖酶(XLN)编码基因(XLN)及其cDNA分别插入载体pVT100,以便在酿酒酵母中表达。表达受其自身启动子或编码乙醇脱氢酶(ADH1)启动子的控制。用含有结构XLN基因cDNA和XLN启动子的质粒转化的酵母,以半乳糖作为碳源生长时会产生有活性的细胞外XLN。然而,以葡萄糖作为碳源时,XLN受到抑制。使用受葡萄糖刺激的ADH1启动子,XLN被分泌到培养基中。在这两种情况下,分泌的48 kDa酶与浅白隐球酵母产生的天然XLN相对应。携带基因组XLN基因的质粒存在转录,但酿酒酵母未将打断XLN的7个内含子剪接出来,也未产生酶。