Bergès T, Barreau C, Peberdy J F, Boddy L M
Laboratoire de Génétique, Université de Bordeaux II URA CNRS 542, Talence, France.
Curr Genet. 1993 Jul-Aug;24(1-2):53-9. doi: 10.1007/BF00324665.
The filamentous fungus Aspergillus niger produces two glycosylated forms of the sucrose-hydrolysing enzyme, invertase. In contrast, some Trichoderma species lack invertase and are unable to utilise sucrose as a sole carbon source. Using an A. niger genomic library constructed in a cosmid vector containing the ura5 gene of Podospora anserina as a selectable marker, and the T. reesei ura5- strain as a sucrose-minus recipient strain, an A. niger invertase gene (suc1) has been cloned by a sib selection procedure. PAGE and enzyme analysis confirmed that transformants had acquired invertase activity. The cloned gene contained DNA sequences which were complementary to the amino-acid sequences of tryptic peptides found in invertase purified from A. niger. The suc1 invertase gene can be used as a dominant selectable marker for the transformation of Trichoderma strains.
丝状真菌黑曲霉产生两种蔗糖水解酶即转化酶的糖基化形式。相比之下，一些木霉属物种缺乏转化酶，无法将蔗糖作为唯一碳源利用。利用构建在含有粪生粪壳菌ura5基因作为选择标记的黏粒载体中的黑曲霉基因组文库，以及里氏木霉ura5-菌株作为蔗糖缺陷型受体菌株，通过 sib 选择程序克隆了黑曲霉转化酶基因（suc1）。聚丙烯酰胺凝胶电泳（PAGE）和酶分析证实转化体获得了转化酶活性。克隆的基因包含与从黑曲霉纯化的转化酶中发现的胰蛋白酶肽氨基酸序列互补的 DNA 序列。suc1 转化酶基因可作为木霉菌株转化的显性选择标记。
