Goldman G H, Vasseur V, Contreras R, Van Montagu M
Laboratorium voor Genetica, Universiteit Gent, Belgium.
Gene. 1994 Jun 24;144(1):113-7. doi: 10.1016/0378-1119(94)90213-5.
The promoters of constitutive genes have proven themselves to be useful parts of expression vectors for genetic engineering in different organisms. Based on a differential screening of an induced cDNA library, we have used a new approach to isolate constitutively expressed genes which can be used for this purpose. Following this strategy, we have isolated constitutively expressed cDNA clones of the mycoparasitic fungus Trichoderma harzianum. One of these cDNA clones corresponds to a gene (cob4) that encodes a novel serine + alanine-rich protein. The predicted amino acid (aa) sequence of this protein (171 aa, deduced M(r) 15,564, pI 12.49) displays a motif that is also found in structural proteins of different species and in a yeast thermoinducible protein. Northern (RNA) blot analysis has demonstrated that cob4 is expressed during growth when glucose or cell walls of a phytopathogenic fungus are provided as carbon source. Hybridizations of T. harzianum genomic DNA indicate that this gene most probably belongs to a multigene family.
组成型基因的启动子已被证明是用于不同生物体基因工程的表达载体的有用组成部分。基于对诱导型cDNA文库的差异筛选,我们采用了一种新方法来分离可用于此目的的组成型表达基因。按照此策略,我们分离出了寄生真菌哈茨木霉组成型表达的cDNA克隆。其中一个cDNA克隆对应于一个基因(cob4),该基因编码一种新型的富含丝氨酸和丙氨酸的蛋白质。该蛋白质的预测氨基酸(aa)序列(171个氨基酸,推导的M(r) 15,564,pI 12.49)显示出一种基序,该基序也存在于不同物种的结构蛋白和酵母热诱导蛋白中。Northern(RNA)印迹分析表明,当以植物病原真菌的葡萄糖或细胞壁作为碳源时,cob4在生长过程中表达。哈茨木霉基因组DNA的杂交表明,该基因很可能属于一个多基因家族。