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Use of the pyrG gene as a food-grade selection marker in Monascus

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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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Acknowledgments

We thank Professor Soo-Un Kim for providing pSGF957. This work was supported by the fund of National Natural Science Foundation of P. R. China (No. 30860123).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanjing East Road 235, Nanchang, Jiangxi, 330047, China

    Bo-hua Wang & Yang Xu

  2. Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang, 330047, Jiangxi, People’s Republic of China

    Yan-ping Li

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  1. Bo-hua Wang
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  2. Yang Xu
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  3. Yan-ping Li
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Correspondence to Yang Xu.

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Wang, Bh., Xu, Y. & Li, Yp. Use of the pyrG gene as a food-grade selection marker in Monascus . Biotechnol Lett 32, 1631–1635 (2010). https://doi.org/10.1007/s10529-010-0336-3

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  • DOI: https://doi.org/10.1007/s10529-010-0336-3

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