Use of the <Emphasis Type="Italic">pyrG</Emphasis> gene as a food-grade selection marker in <Emphasis Type="Italic">Monascus</Emphasis> |
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Authors: | Bo-hua Wang Yang Xu Yan-ping Li |
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Institution: | (1) State Key Laboratory of Food Science and Technology, Nanchang University, Nanjing East Road 235, Nanchang, Jiangxi, 330047, China;(2) Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, 330047, Jiangxi, People’s Republic of China; |
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Abstract: | Ma-pyrG was cloned from Monascus aurantiacus AS3.4384 using degenerate PCR with primers designed with an algorithm called CODEHOP, and its complete sequence was obtained
by a PCR-based strategy for screening a Monascus fosmid library. Ma-pyrG encodes orotidine-5′-phosphate decarboxylase (OMPdecase), a 283-aminoacid protein with 81% sequence identity to that from
Aspergillus flavus NRRL 3357. A pyrG mutant strain from M. aurantiacus AS3.4384, named UM28, was isolated by resistance to 5-fluoroorotic acid after UV mutagenesis. Sequence analysis of this mutated
gene revealed that it contained a point mutation at nucleotide position +220. Plasmid pGFP-pyrG, bearing the green fluorescent protein gene (GFP) as a model gene and Ma-pyrG as a selection marker, were constructed. pGFP-pyrG were successfully transformed into UM28 by using the PEG method. |
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