TRYPTOPHAN ACCUMULATION IN SACCHAROMYCES-CEREVISIAE UNDER THE INFLUENCE OF AN ARTIFICIAL YEAST TRP GENE-CLUSTER
TRYPTOPHAN ACCUMULATION IN SACCHAROMYCES-CEREVISIAE UNDER THE INFLUENCE OF AN ARTIFICIAL YEAST TRP GENE-CLUSTER
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DOI：
10.1002/yea.320030206
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发表时间：
1987-06-01
期刊：
影响因子：
2.6
通讯作者：
HUTTER, R
中科院分区：
文献类型：
--
作者：
PRASAD, R;NIEDERBERGER, P;HUTTER, R
构建了携带所有5个酵母TRP基因的质粒pME 559。该质粒是基于pBR 322和源自质粒pJDB 207的2 μ m-DNA序列的酵母/大肠杆菌穿梭载体。我们在酵母中研究了（i）在选择性和非选择性条件下质粒的稳定性，（ii）所有五种色氨酸基因的表达和（iii）酵母转化体中色氨酸的积累。这些研究是与早期的构建体pME 554进行比较的，pME 554在TRP 1基因的表达方面与质粒pME 559不同，并且携带TRP 2野生型而不是TRP 2fbr突变等位基因。为了在酵母中稳定维持质粒，选择是必要的。质粒pME 559显示所有色氨酸基因的正常表达，并且发现酶水平平均比野生型菌株高23倍。相比之下，在这样的质粒携带菌株中观察到的最大色氨酸通量比野生型菌株中的最大通量能力高约10倍。
Plasmid pME559, carrying all five yeastTRPgenes, was constructed. This plasmid is a yeast/Escherichia colishuttle vector based on pBR322 and 2 μm‐DNA sequences derived from plasmid pJDB207. We studied in yeast (i) the stability of the plasmid under selective and non‐selective conditions, (ii) expression of all fiveTRPgenes and (iii) tryptophan accumulation in yeast transformants. These studies were conducted in comparison with an earlier construction, pME554, which differs from plasmid pME559 in the expression of theTRP1gene and which carries theTRP2wild type instead of theTRP2fbrmutant allele. For stable maintenance of the plasmids in yeast a selection was necessary. Plasmid pME559 displayed normal expression of allTRPgenes, and enzyme levels on average 23‐fold higher than in the wild type strain were found. In comparison, the maximal tryptophan flux observed in such a plasmid‐carrying strain was about ten‐fold higher than the maximal flux capacity in the wild type strain.