The application of PCR for the isolation of a lipase gene from the genomic DNA of an Antarctic microfungus

We successfully isolated a lipase gene (designated lipPA) directly from the genomic DNA of an Antarctic isolate of Penicillium allii using PCR and a suite of degenerate primers specifically designed to target two conserved regions of fungal lipase genes. We applied the biolistic transformation syste...

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Bibliographic Details
Published in:Current Genetics
Main Authors: Bradner, J. Ron, Bell, Philip J L, Te'o, V. S Junior, Nevalainen, K. M Helena
Format: Article in Journal/Newspaper
Language:English
Published: 2003
Subjects:
Antarctic
Antarc*
Online Access:https://researchers.mq.edu.au/en/publications/0f90bbbd-5aa5-4a78-aa64-0c267ea66d45
https://doi.org/10.1007/s00294-003-0440-1
http://www.scopus.com/inward/record.url?scp=0347513481&partnerID=8YFLogxK
Description
Summary:We successfully isolated a lipase gene (designated lipPA) directly from the genomic DNA of an Antarctic isolate of Penicillium allii using PCR and a suite of degenerate primers specifically designed to target two conserved regions of fungal lipase genes. We applied the biolistic transformation system to successfully integrate the lipPA gene into a heterologous fungal host, Trichoderma reesei, one of the most powerful secretors of extracellular proteins, and induced the transformant to secrete an active lipase into the growth medium. The recombinant lipase had a temperature optimum of 25 °C at pH 7.9 and retained greater than 50% of the maximum activity from 10 °C to 35 °C and over a pH range from 4.0 to 8.5.

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