Abstrak 
Transformasi Plasmid pUR5750 Ke Dalam Sel Monascuz Purpures ITBCC-HD-F002 Melalui Mediasi Bakteri Agrobacterium Tumefaciens LBA1100 Dan Agl1
Tiana Milanda , Syarif Hamdani, Marlia S. Wibowo, Tutus Gusdinar, Haryanto Dhanutirto, Philippe J. Blanc
Universitas Padjadjaran, Seminar Nasional “ Reposisi Peran Pendidikan Tinggi Farmasi Dalam Penyediaan Sumber Daya Manusia dan Hasil Riset Bagi Industri Nasional” Dalam Rangka Peringatan 50 Tahun Pendidikan Jasmani Universitas Farmasi Bandung, 20 Maret 2009
Bahasa Indonesia, Bahasa Inggris
Universitas Padjadjaran, Seminar Nasional “ Reposisi Peran Pendidikan Tinggi Farmasi Dalam Penyediaan Sumber Daya Manusia dan Hasil Riset Bagi Industri Nasional” Dalam Rangka Peringatan 50 Tahun Pendidikan Jasmani Universitas Farmasi Bandung, 20 Maret 2009
Agrobacterium tumefaciens, Monascus purpureus, pUR5750, transformation
Monascus purpureus is a red fungus that produces various secondary metabolites, such as pigments, monacolin K, and monascidin A. The pigments are traditionally used as food and cosmetic colorants, whereas monacolin K shows an antihypercholesterolemic activity. Monascidin A shows an antimicrobial activity, but it has carcinogenic and nephrotoxic properties. Many researches have been done to eliminate this toxic metabolite. Since monascidin A’s precursor is similar to its pigments, our research was aimed to develop an efficient genetic transformation system that can be used to identify monascidin A genes. The transformation must be applied in a nonproducing pigment (albino) mutant. M purpureus ITBCC-HD-F002, an EMS-induced albino mutant, was transformed into hygromycin B resistance using the hygromycin B phosphotransferase (hph) of Escherichia colt as the selective trait, governed by gdp promoter of Aspergillus nidulans in pUR5750 plasmid. This plasmid was transformed into spores or protoplasts from this fungus mediated by Agrobacterium LBA1100 and AGL1. The transformation efficiency was 346 ± 4.00 transformants/107 protoplast (mediated by A. tumefaciens LBA11) and 635.7 ± 13.32 transformants/107 spores (mediated by A. tumefaciens AGL1). These results indicating the highly virulent strain A. tumefaciens AGL1 was found to be more efficient in DNA transfer than LBA1100. The majority of transformants were mitotically stable up to five generations and the presence of hph genes were detected by PCR. In four randomly chosen transformants, single-copy integration of the marker gene at different chromosomal site were proven by Southern Blot analysis.