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

J. Ron Bradner; Philip J L Bell; V. S Junior Te'o; K. M Helena Nevalainen

doi:10.1007/s00294-003-0440-1

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

J. Ron Bradner^*, Philip J L Bell, V. S Junior Te'o, K. M Helena Nevalainen

^*Corresponding author for this work

Faculty of Science and Engineering

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	224-230
Number of pages	7
Journal	Current Genetics
Volume	44
Issue number	4
DOIs	https://doi.org/10.1007/s00294-003-0440-1
Publication status	Published - Dec 2003

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abstract = "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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AB - 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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The application of PCR for the isolation of a lipase gene from the genomic DNA of an Antarctic microfungus

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