Cloning of an Aspergillus niger invertase gene by expression in Trichoderma reesei.
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.
Bergès
T
T
Laboratoire de Génétique, Université de Bordeaux II URA CNRS 542, Talence, France.
Barreau
C
C
Peberdy
J F
JF
Boddy
L M
LM
D001234:Aspergillus niger / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D003360:Cosmids; D015723:Gene Library; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D005819:Genetic Markers; D006026:Glycoside Hydrolases / Q000096:biosynthesis / Q000235:genetics*; D015183:Restriction Mapping; D013395:Sucrose / Q000378:metabolism; D014170:Transformation, Genetic; D014242:Trichoderma / Q000235:genetics* / Q000254:growth & development; D043324:beta-Fructofuranosidase
D005819:Genetic Markers; D013395:Sucrose; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase