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Expression of the lacZ gene from two methanol-regulated promoters in Pichia pastoris

机译:来自两个甲醇调节启动子的lacZ基因在巴斯德毕赤酵母中的表达

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Two DNA fragments containing putative control regions regulating the expression of the alcohol oxidase (AOX) and dihydroxy-acetone synthase (DAS) genes from the methylotrophic yeast Pichia pastoris were used in the construction of vectors for the expression of the Escherichia coli lacZ gene. These vectors were transformed into P. pastoris host cells and employed in experiments to measure the control mechanisms employed by each promoter in the production of β-galactosidase fusion products. Results in P. pastoris suggest that the processes used to regulate the expression of these gene fusions involve both repression/dere-pression and induction mechanisms. Expression of the AOX-lacZ and DAS-lacZ fusions was examined in Saccharomyces cerevisiae as well. Interestingly, β-galactosidase was expressed in a regulated manner in the heterologous host.
机译:使用两个含有推测的控制区的DNA片段,这些控制区调节了来自甲基营养酵母巴斯德毕赤酵母的醇氧化酶(AOX)和二羟基丙酮合酶(DAS)基因的表达,用于构建表达大肠杆菌lacZ基因的载体。将这些载体转化到巴斯德毕赤酵母宿主细胞中,并用于实验中以测量每个启动子在生产β-半乳糖苷酶融合产物中所采用的控制机制。巴斯德毕赤酵母的结果表明,用于调节这些基因融合体表达的过程涉及阻抑/抑制和诱导机制。也在酿酒酵母中检查了AOX-lacZ和DAS-lacZ融合体的表达。有趣的是,β-半乳糖苷酶在异源宿主中以受控方式表达。

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