Publication: Development of transformation system in Monascus purpureus using an autonomous replication vector with aureobasidin A resistance gene
Issued Date
2006-01-01
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ISSN
15736776
01415492
01415492
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2-s2.0-29244479966
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Mahidol University
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SCOPUS
Bibliographic Citation
Biotechnology Letters. Vol.28, No.2 (2006), 115-120
Suggested Citation
Takeo Shimizu, Hiroshi Kinoshita, Takuya Nihira Development of transformation system in Monascus purpureus using an autonomous replication vector with aureobasidin A resistance gene. Biotechnology Letters. Vol.28, No.2 (2006), 115-120. doi:10.1007/s10529-005-4956-y Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/23105
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Title
Development of transformation system in Monascus purpureus using an autonomous replication vector with aureobasidin A resistance gene
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Abstract
To enhance the variety of genetic tools and thus to promote molecular genetic study, aureobasidin A and its resistance gene were adopted as a new marker system together with the incorporation of the Gateway system to facilitate the introduction of long heterologous DNA fragments into Monascus purpureus. The minimum inhibitory concentration of aureobasidin A against Monascus was 0.05 μg/ml and a transformation efficiency of 17 colonies/μg DNA was obtained by the protoplast-PEG method with the vector pAUR316, containing the aureobasidin A resistance gene. Southern analysis of the transformants confirmed that pAUR316 exists as an independent vector, demonstrating that the AMA1 sequence acts as the autonomous replication sequence in M. purpureus. Through the use of the Gateway system, a polyketide synthase gene (7.8 kbp) responsible for citrinin biosynthesis was introduced. As a result, the transformants showed 1.5-fold higher production of citrinin than the wild-type strain. © Springer 2006.