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[Construct cosmid libraries by isolating large genomic DNA fragments from Monascus ruber]. 2009

Yang An, and Jing Yang, and Xinxin Xu, and Gang Liu
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. anyang@foxmail.com

OBJECTIVE To isolate large genomic DNA fragments from Monascus ruber for the construction of cosmid libraries. METHODS Modified phenol-chloroform method was used to isolate genomic DNA. The isolated genomic DNA was digested by Sau3AI to 40kb fragments on average. Then, the fragments were packaged by Stratagene' s Gigapack III XL packaging extract. A pair of degenerate primers were used to amplify a fragment of PKS (polyketide synthase) gene from this cosmid library. RESULTS The average size of genomic DNA isolated by this method was larger than 48 kb, with a concentration of 5 microg/microl. The constructed cosmid libraries had 10 folders coverage of the Monascus spp. genome. A cosmid containing the homologue of PKS gene was obtained by PCR screening. CONCLUSIONS This modified method of isolating large fragments genomic DNA of Monascus ruber was efficient and feasible.

UI MeSH Term Description Entries
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D003360 Cosmids Plasmids containing at least one cos (cohesive-end site) of PHAGE LAMBDA. They are used as cloning vehicles. Cosmid
D004271 DNA, Fungal Deoxyribonucleic acid that makes up the genetic material of fungi. Fungal DNA
D005821 Genetic Techniques Chromosomal, biochemical, intracellular, and other methods used in the study of genetics. Genetic Technic,Genetic Technics,Genetic Technique,Technic, Genetic,Technics, Genetic,Technique, Genetic,Techniques, Genetic
D015723 Gene Library A large collection of DNA fragments cloned (CLONING, MOLECULAR) from a given organism, tissue, organ, or cell type. It may contain complete genomic sequences (GENOMIC LIBRARY) or complementary DNA sequences, the latter being formed from messenger RNA and lacking intron sequences. DNA Library,cDNA Library,DNA Libraries,Gene Libraries,Libraries, DNA,Libraries, Gene,Libraries, cDNA,Library, DNA,Library, Gene,Library, cDNA,cDNA Libraries
D032741 Monascus A genus in the family Monascaceae, order EUROTIALES. One species, Monascus purpureus, has multiple uses in traditional Chinese medicine (MEDICINE, CHINESE TRADITIONAL). Monascus albidus,Monascus anka,Monascus araneosus,Monascus rubiginosus,Monascus purpureus

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