|Skinner, Daniel - Dan|
Submitted to: Electronic Journal of Biotechnology
Publication Type: Peer reviewed journal
Publication Acceptance Date: 5/22/2003
Publication Date: 8/15/2003
Citation: Lakrod, K., Chaisrisook, C., Skinner, D.Z. 2003. Transformation of monascus purpureus to hygromycin b resistance with cosmid pmocosx reduces fertility. Electronic Journal of Biotechnology. http://www.ejbiotechnology.info/content/vol6/issue2/full/3/index.html Interpretive Summary: The fungus Monascus purpureus, also known as the ¿Red Rice¿ fungus, is used in the production of various Asian food products. This study was conducted to identify parameters important to successful genetic transformation of this fungus in order to facilitate future studies of various genes. Using high-pressure helium and microscopic tungsten particles coated with a gene conditioning fungicide resistance, the fungus was transformed to fungicide resistance. The resulting strains were severely restricted in their ability to sexually reproduce, suggesting the expression of the fungicide resistance gene interfered with the normal reproduction processes.
Technical Abstract: Albino strain KB20M1 of Monascus purpureus was genetically transformed to hygromycin B resistance with cosmid pMOcosX, using biolistic bombardment. Conidia and mycelial fragments were used as the recipient material and rupture disk strengths of 6.2 and 9.3 MPa each yielded a transformed isolate from five bombardments. Southern analysis suggested a single copy of the cosmid had integrated into each of the transformants. Both of the independent transformants formed cleistothecia but ascospore formation was greatly reduced or absent, suggesting the integration and expression of the genes carried on pMOcosX interfered with fertilization and/or ascospore formation in this homothallic fungus.