Transformation System for Hypocrea jecorina (Trichoderma reesei) That Favors Homologous Integration and Employs Reusable Bidirectionally Selectable Markers
Matthias G. Steiger,Marika Vitikainen,Pekka Uskonen,Kurt Brunner,Gerhard Adam,Tiina Pakula,Merja Penttilä,Markku Saloheimo,Robert L. Mach,Astrid R. Mach-Aigner +9 more
TLDR
A transformation system that allows highly efficient gene targeting by using a tmus53 (human LIG4 homolog) deletion strain and permits the unlimited reuse of the same marker by employing a Cre/loxP-based excision system is reported.Abstract:
Hypocrea jecorina is an industrially important filamentous fungus due to its effective production of hydrolytic enzymes. It has received increasing interest because of its ability to convert lignocellulosic biomass to monomeric sugars, which can be converted into biofuels or platform chemicals. Genetic engineering of strains is a highly important means of meeting the requirements of tailor-made applications. Therefore, we report the development of a transformation system that allows highly efficient gene targeting by using a tmus53 (human LIG4 homolog) deletion strain. Moreover, it permits the unlimited reuse of the same marker by employing a Cre/loxP-based excision system. Both marker insertion and marker excision can be positively selected for by combining resistance to hygromycin B and loss of sensitivity to fluoroacetamide. Finally, the marker pyr4, also positively selectable for insertion and loss, can be used to remove the cre gene.read more
Citations
More filters
Journal ArticleDOI
A versatile toolkit for high throughput functional genomics with Trichoderma reesei
André Schuster,André Schuster,Kenneth S. Bruno,James R. Collett,Scott E. Baker,Bernhard Seiboth,Christian P. Kubicek,Monika Schmoll +7 more
TL;DR: A construction kit for gene knock out in T. reesei and a primer database for gene deletion using the pyr4, amdS and hph selection markers are provided to boost research towards efficient exploitation of the resources of T.Reesei for cellulase expression and hence second generation biofuel production.
Journal ArticleDOI
Trichoderma reesei RUT-C30--thirty years of strain improvement.
Robyn Peterson,Helena Nevalainen +1 more
TL;DR: The results from biochemical-, microscopic-, genomic-, transcriptomic-, glycomic- and proteomic-based research on the RUT-C30 strain published over the last 30 years are discussed.
Journal ArticleDOI
Cellulases and beyond: the first 70 years of the enzyme producer Trichoderma reesei.
TL;DR: This review summarizes important milestones of the development of T. reesei as the leading production host for biorefinery enzymes, and discusses emerging trends in strain engineering of the filamentous ascomycete TrichodermaReesei.
Journal ArticleDOI
Development of a low-cost cellulase production process using Trichoderma reesei for Brazilian biorefineries
Simo Ellilä,Lucas Miranda Fonseca,Cristiane Uchima,Junio Cota,Gustavo H. Goldman,Markku Saloheimo,Vera Sacon,Matti Siika-aho +7 more
TL;DR: It is shown that with relatively few genetic modifications and the right growth medium it is possible to produce considerable amounts of well-performing cellulase at very low cost in Brazil using T. reesei.
Journal ArticleDOI
Genetic engineering of Trichoderma reesei cellulases and their production
TL;DR: The structure and mechanism of the enzymes involved, the regulation of their expression and the pathways of their formation and secretion have been investigated in T. reesei in considerable details and several of the findings have been used to improve the formation of the T.Reesei cellulases and their properties.
References
More filters
Journal ArticleDOI
The path forward for biofuels and biomaterials
Arthur J. Ragauskas,Charlotte K. Williams,Brian H. Davison,George J. P. Britovsek,John Cairney,Charles A. Eckert,William J. Frederick,Jason P. Hallett,David J. Leak,Charles L. Liotta,Jonathan R. Mielenz,Richard J. Murphy,Richard H. Templer,Timothy J. Tschaplinski +13 more
TL;DR: The integration of agroenergy crops and biorefinery manufacturing technologies offers the potential for the development of sustainable biopower and biomaterials that will lead to a new manufacturing paradigm.
Journal ArticleDOI
Biomass recalcitrance: engineering plants and enzymes for biofuels production.
Michael E. Himmel,Shi You Ding,David K. Johnson,William S. Adney,Mark R. Nimlos,John W. Brady,Thomas D. Foust +6 more
TL;DR: Here, the natural resistance of plant cell walls to microbial and enzymatic deconstruction is considered, collectively known as “biomass recalcitrance,” which is largely responsible for the high cost of lignocellulose conversion.
Journal ArticleDOI
A New Efficient Gene Disruption Cassette for Repeated Use in Budding Yeast
TL;DR: A loxP-kanMX-loxP gene disruption cassette is developed, which combines the advantages of the heterologous kanr marker with those from the Cre- lox P recombination system, and will be of great advantage for the functional analysis of gene families.
Journal ArticleDOI
Multifunctional yeast high-copy-number shuttle vectors
TL;DR: A set of four yeast shuttle vectors that incorporate sequences from the Saccharomyces cerevisiae 2 mu endogenous plasmid has been constructed, providing high-copy-number counterparts to the current pRS vectors.
Journal ArticleDOI
Bio-ethanol--the fuel of tomorrow from the residues of today.
TL;DR: This review gives an overview of the new technologies required and the advances achieved in recent years to bring lignocellulosic ethanol towards industrial production.
Related Papers (5)
Genome sequencing and analysis of the biomass-degrading fungus Trichoderma reesei (syn. Hypocrea jecorina).
Diego Martinez,Diego Martinez,Randy M. Berka,Bernard Henrissat,Markku Saloheimo,Mikko Arvas,Scott E. Baker,Jarod Chapman,Olga Chertkov,Pedro M. Coutinho,Dan Cullen,Etienne Danchin,Igor V. Grigoriev,Paul Harris,Melissa Jackson,Christian P. Kubicek,Cliff Han,Isaac Ho,Luis F. Larrondo,Alfredo Lopez de Leon,Jon K. Magnuson,Sandy Merino,Monica Misra,Beth Nelson,Nicholas H. Putnam,Barbara Robbertse,Asaf Salamov,Monika Schmoll,Astrid Terry,Nina Thayer,Ann Westerholm-Parvinen,Conrad L. Schoch,Jian Yao,Ravi D. Barabote,Mary Anne Nelson,Chris Detter,David Bruce,Cheryl R. Kuske,Gary Xie,Paul G. Richardson,Daniel S. Rokhsar,Susan Lucas,Edward M. Rubin,Nigel Dunn-Coleman,Michael Ward,Thomas Brettin +45 more