Transformation System for Hypocrea jecorina ( Trichoderma reesei ) That Favors Homologous Integration and Employs Reusable Bidirectionally Selectable Markers

• DOI: 10.1128/aem.02100-10
• Publication Date: 2010-11-13T05:59:41Z
• Authors: Matthias G. Steiger; Marika Vitikainen; Pekka Uskonen; Kurt Brunner; Gerhard Adam; Tiina Pakula; Merja Penttilä; Markku Saloheimo; Robert L. Mach; Astrid R. Mach-Ai...
• Keywords: Hypocrea; Trichoderma; Selectable marker

ABSTRACT Hypocrea jecorina is an industrially important filamentous fungus due to its effective production of hydrolytic enzymes. It has received increasing interest because ability convert lignocellulosic biomass monomeric sugars, which can be converted into biofuels or platform chemicals. Genetic engineering strains a highly means meeting the requirements tailor-made applications. Therefore, we report development transformation system that allows efficient gene targeting by using tmus53 (human LIG4 homolog) deletion strain. Moreover, it permits unlimited reuse same marker employing Cre/loxP-based excision system. Both insertion and positively selected for combining resistance hygromycin B loss sensitivity fluoroacetamide. Finally, pyr4 , also selectable loss, used remove cre gene.