Promoter-tagged restriction enzyme-mediated insertion (PT-REMI) mutagenesis in Aspergillus niger

Shuster, J. R.; Connelley, M. Bindel

doi:10.1007/pl00008638

Cited by 12 publications

(13 citation statements)

References 36 publications

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“…The ability to introduce or delete genes remains difficult although some advances in transformation have been reported, e.g., restriction enzyme-mediated integration [34] and Agrobacterium tumefaciens-Ti plasmid mediated transformation [35]. Levels of production of non-fungal proteins have been low compared to those of homologous proteins.…”

Section: Filamentous Fungimentioning

confidence: 98%

Contributions of Microorganisms to Industrial Biology

2007

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Life on earth is not possible without microorganisms. Microbes have contributed to industrial science for over 100 years. They have given us diversity in enzymatic content and metabolic pathways. The advent of recombinant DNA brought many changes to industrial microbiology. New expression systems have been developed, biosynthetic pathways have been modified by metabolic engineering to give new metabolites, and directed evolution has provided enzymes with modified selectability, improved catalytic activity and stability. More and more genomes of industrial microorganisms are being sequenced giving valuable information about the genetic and enzymatic makeup of these valuable forms of life. Major tools such as functional genomics, proteomics, and metabolomics are being exploited for the discovery of new valuable small molecules for medicine and enzymes for catalysis.

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Section: Filamentous Fungimentioning

confidence: 98%

Contributions of Microorganisms to Industrial Biology

2007

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“…(Kuspa and Loomis 1992), Aspergillus sp. (Shuster and Bindel 1999), Ustilago maydis (Bolker et al 1995), Coprinus cinereus (Granado et al 1997), Lentinus edodes (Sato et al 1998), Polysphondylium pallidum (Kawabe et al 1999), Gibberella fujikuroi (Linnemannstons et al 1999) and Candida albicans (Brown et al 1996).…”

Section: Introductionmentioning

confidence: 98%

Tagging Hansenula polymorpha genes by random integration of linear DNA fragments (RALF)

et al. 2001

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We have investigated the feasibility of using gene tagging by restriction enzyme-mediated integration (REMI) to isolate mutants in Hansenula polymorpha. A plasmid that cannot replicate in H. polymorpha and contains a dominant zeocin resistance cassette, pREMI-Z, was used as the integrative/mutagenic plasmid. We observed that high transformation efficiency was primarily dependent on the use of linearised pREMI-Z, and that the addition of restriction endonuclease to linearised pREMI-Z prior to transformation increased the transformation frequency only slightly. Integration of linearised pREMI-Z occurred at random in the H. polymorpha genome. Therefore, we termed this method Random integration of Linear DNA Fragments (RALF). To explore the potential of RALF in H. polymorpha, we screened a collection of pREMI-Z transformants for mutants affected in peroxisome biogenesis (pex) or selective peroxisome degradation (pdd). Many previously described PEX genes were obtained from the mutant collection, as well as a number of new genes, including H. polymorpha PEX12 and genes whose function in peroxisome biogenesis is still unclear. These results demonstrate that RALF is a powerful tool for tagging genes in H. polymorpha that should make it possible to carry out genome-wide mutagenesis screens.

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“…Promoter-tagged restriction enzyme-mediated insertion (PT-REMI) is a variant of REMI, which was developed in Aspergillus niger (Shuster and Connelley 1999 ). A vector is used that contains a strong promoter.…”

Section: Historical and Modernmentioning

confidence: 99%