Kupper U, Linden M, Cao K Z, Lerch K
Biochemisches Institut der Universität Zürich, Switzerland.
Curr Genet. 1990 Nov;18(4):331-5. doi: 10.1007/BF00318214.
Wild-type Neurospora crassa, strain Singapore, was transformed with a N. crassa metallothionein promoter/protyrosinase fusion gene. Transformants produced tyrosinase during vegetative growth, as determined by Western analyses and activity assays. This is in sharp contrast to wild-type strains, where this enzyme is only expressed in situations of starvation or sexual differentiation. Complete integration of a 400 bp metallothionein promoter-fragment leads to constitutive expression of protyrosinase, whereas a 3.6 kb promoter-fragment conferred copper inducibility on the reporter gene in four transformants. A transformant with high constitutive tyrosinase levels was able to produce melanin on complete medium agar plates supplemented with 1 mg/ml L-tyrosine.
用粗糙脉孢菌金属硫蛋白启动子/原酪氨酸酶融合基因转化野生型粗糙脉孢菌新加坡菌株。通过蛋白质免疫印迹分析和活性测定确定，转化体在营养生长期间产生酪氨酸酶。这与野生型菌株形成鲜明对比，在野生型菌株中，这种酶仅在饥饿或有性分化情况下表达。400 bp金属硫蛋白启动子片段的完全整合导致原酪氨酸酶的组成型表达，而在四个转化体中，一个3.6 kb的启动子片段赋予报告基因铜诱导性。一个具有高组成型酪氨酸酶水平的转化体能够在补充有1 mg/ml L-酪氨酸的完全培养基琼脂平板上产生黑色素。
