DNA Assembler Method for Construction of Zeaxanthin-producing Strains of Saccharomyces Cerevisiae

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2012 Jun 20

PMID 22711131

Citations 11

Authors

Zengyi Shao

Yunzi Luo

Huimin Zhao

Affiliations

Soon will be listed here.

Abstract

DNA assembler enables design and rapid construction of biochemical pathways in a one-step fashion by exploitation of the in vivo homologous recombination mechanism in Saccharomyces cerevisiae. It has many applications in pathway engineering, metabolic engineering, combinatorial biology, and synthetic biology. Here we use the zeaxanthin biosynthetic pathway as an example to describe the key steps in the construction of pathways containing multiple genes using the DNA assembler approach. Methods for the construction of the clones, S. cerevisiae transformation, and zeaxanthin production and detection are shown.

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References

Hjersted J, Henson M, Mahadevan R . Genome-scale analysis of Saccharomyces cerevisiae metabolism and ethanol production in fed-batch culture. Biotechnol Bioeng. 2007; 97(5):1190-204. DOI: 10.1002/bit.21332. View

Szczebara F, Chandelier C, Villeret C, Masurel A, Bourot S, Duport C . Total biosynthesis of hydrocortisone from a simple carbon source in yeast. Nat Biotechnol. 2003; 21(2):143-9. DOI: 10.1038/nbt775. View

Chemler J, Yan Y, Koffas M . Biosynthesis of isoprenoids, polyunsaturated fatty acids and flavonoids in Saccharomyces cerevisiae. Microb Cell Fact. 2006; 5:20. PMC: 1533850. DOI: 10.1186/1475-2859-5-20. View

Misawa N, Nakagawa M, Kobayashi K, Yamano S, Izawa Y, Nakamura K . Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. J Bacteriol. 1990; 172(12):6704-12. PMC: 210783. DOI: 10.1128/jb.172.12.6704-6712.1990. View

Gunyuzlu P, Hollis G, Toyn J . Plasmid construction by linker-assisted homologous recombination in yeast. Biotechniques. 2002; 31(6):1246, 1248, 1250. DOI: 10.2144/01316bm03. View

Yan Y, Kohli A, Koffas M . Biosynthesis of natural flavanones in Saccharomyces cerevisiae. Appl Environ Microbiol. 2005; 71(9):5610-3. PMC: 1214632. DOI: 10.1128/AEM.71.9.5610-5613.2005. View

Oldenburg K, Vo K, Michaelis S, Paddon C . Recombination-mediated PCR-directed plasmid construction in vivo in yeast. Nucleic Acids Res. 1997; 25(2):451-2. PMC: 146432. DOI: 10.1093/nar/25.2.451. View

Ma H, Kunes S, Schatz P, Botstein D . Plasmid construction by homologous recombination in yeast. Gene. 1987; 58(2-3):201-16. DOI: 10.1016/0378-1119(87)90376-3. View

Pitera D, Paddon C, Newman J, Keasling J . Balancing a heterologous mevalonate pathway for improved isoprenoid production in Escherichia coli. Metab Eng. 2007; 9(2):193-207. DOI: 10.1016/j.ymben.2006.11.002. View

10.

Lee F, da Silva N . Sequential delta-integration for the regulated insertion of cloned genes in Saccharomyces cerevisiae. Biotechnol Prog. 1997; 13(4):368-73. DOI: 10.1021/bp970055d. View

11.

Shao Z, Zhao H, Zhao H . DNA assembler, an in vivo genetic method for rapid construction of biochemical pathways. Nucleic Acids Res. 2008; 37(2):e16. PMC: 2632897. DOI: 10.1093/nar/gkn991. View

12.

Misawa N, Shimada H . Metabolic engineering for the production of carotenoids in non-carotenogenic bacteria and yeasts. J Biotechnol. 1997; 59(3):169-81. DOI: 10.1016/s0168-1656(97)00154-5. View

13.

Ro D, Paradise E, Ouellet M, Fisher K, Newman K, Ndungu J . Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Nature. 2006; 440(7086):940-3. DOI: 10.1038/nature04640. View

14.

Dejong J, Liu Y, Bollon A, Long R, Jennewein S, Williams D . Genetic engineering of taxol biosynthetic genes in Saccharomyces cerevisiae. Biotechnol Bioeng. 2005; 93(2):212-24. DOI: 10.1002/bit.20694. View

15.

Horton R, Hunt H, Ho S, Pullen J, Pease L . Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989; 77(1):61-8. DOI: 10.1016/0378-1119(89)90359-4. View

16.

Keasling J . Synthetic biology for synthetic chemistry. ACS Chem Biol. 2008; 3(1):64-76. DOI: 10.1021/cb7002434. View

17.

Raymond C, Pownder T, Sexson S . General method for plasmid construction using homologous recombination. Biotechniques. 1999; 26(1):134-8, 140-1. DOI: 10.2144/99261rr02. View

18.

Menzella H, Reid R, Carney J, Chandran S, Reisinger S, Patel K . Combinatorial polyketide biosynthesis by de novo design and rearrangement of modular polyketide synthase genes. Nat Biotechnol. 2005; 23(9):1171-6. DOI: 10.1038/nbt1128. View