[Construct cosmid libraries by isolating large genomic DNA fragments from Monascus ruber].
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.
An
Yang
Y
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China. anyang@foxmail.com
Yang
Jing
J
Xu
Xinxin
X
Liu
Gang
G
D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics / Q000302:isolation & purification*; D015723:Gene Library; D005821:Genetic Techniques; D032741:Monascus / Q000235:genetics*
D004271:DNA, Fungal