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Content archived on 2024-04-15

DEVELOPMENT OF A GENE MEDIATED TRANSFER AND SELECTION SYSTEM FOR FILAMENTOUS FUNGI

Objective


A method has been developed to genetically transform the fungus Gibberella fujikuroi with exogenous deoxyribonucleic acid (DNA). We have achieved the genetic transformation of a Gibberella mutant using a gene from another fungus of biotechnological interest, Aspergillus nidulans. We have also isolated mutants of the fungus Gibberella fujikuroi that produce mixtures of gibberellins and their precursors which are different from those of the wild type. Simplified methods have been developed for the measurement of gibberellin concentrations and the screening of cultures of the fungus Gibberella fujikuroi with possible alterations in gibberellin production. Combination of these methods with our methods for the isolation of mutants resulted in the isolation of 12 new mutants of Gibberella with alterations in gibberellin biosynthesis.

A study was made fundamental and applied geneticaspects of filamentous fungi involving the development of genetic transformation and expression systems which are of central importance for the study and exploitation of filamentous fungi. Studies focussed on 2 selection systems which have a concomitant phenotype (ie, resistance to an antibiotic agent resulting in a specific growth defect). Certain acceptable fungi were programmed to synthesize and secrete heterologous proteins of commercial value in copious amounts.

Transformation systems were successfully generated for a variety of industrial fungi based on the nitrate reductase system (niaD). These include Aspergillus oryzae, A niger, Penicillium chrysogenum, Gibberella fujikuroi, Cephalosporium acremonium. Transformation based on the uracil system (pyrG) was also developed for A oryzae and A niger. Such results indicate that 2 reproducible reliable routes exist that introduce genes of commercial interest into filamentous fungi.
Deoxyribonucleic acid (DNA) constructs were also made in order to genetically programme fungi to express genes of commercial interest. Initial investigations focussed on the expression and secretion of human interleukin-6 (hIL-6). Such studies resulted in the production of this substance by filamentous fungi.

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Coordinator

UNIV. DE SEVILLA
EU contribution
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Address
RECTORADO C/SAN FERNANDO 4
41071 SEVILLA
Spain

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