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One-step integration of multiple genes into the oleaginous yeast Yarrowia lipolytica. 2014

Shuliang Gao, and Linna Han, and Li Zhu, and Mei Ge, and Sheng Yang, and Yu Jiang, and Daijie Chen
College of Bioengineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China, slgao@cibt.ac.cn.

Yarrowia lipolytica is an unconventional yeast, and is generally recognized as safe (GRAS). It provides a versatile fermentation platform that is used commercially to produce many added-value products. Here we report a multiple fragment assembly method that allows one-step integration of an entire β-carotene biosynthesis pathway (~11 kb, consisting of four genes) via in vivo homologous recombination into the rDNA locus of the Y. lipolytica chromosome. The highest efficiency was 21%, and the highest production of β-carotene was 2.2 ± 0.3 mg per g dry cell weight. The total procedure was completed in less than one week, as compared to a previously reported sequential gene integration method that required n weeks for n genes. This time-saving method will facilitate synthetic biology, metabolic engineering and functional genomics studies of Y. lipolytica.

UI MeSH Term Description Entries
D008967 Molecular Biology A discipline concerned with studying biological phenomena in terms of the chemical and physical interactions of molecules. Biochemical Genetics,Biology, Molecular,Genetics, Biochemical,Genetics, Molecular,Molecular Genetics,Biochemical Genetic,Genetic, Biochemical,Genetic, Molecular,Molecular Genetic
D004271 DNA, Fungal Deoxyribonucleic acid that makes up the genetic material of fungi. Fungal DNA
D004275 DNA, Ribosomal DNA sequences encoding RIBOSOMAL RNA and the segments of DNA separating the individual ribosomal RNA genes, referred to as RIBOSOMAL SPACER DNA. Ribosomal DNA,rDNA
D005827 Genetics, Microbial A subdiscipline of genetics which deals with the genetic mechanisms and processes of microorganisms. Microbial Genetics,Genetic, Microbial,Microbial Genetic
D013997 Time Factors Elements of limited time intervals, contributing to particular results or situations. Time Series,Factor, Time,Time Factor
D053898 Biosynthetic Pathways Sets of enzymatic reactions occurring in organisms and that form biochemicals by making new covalent bonds. Biosynthetic Pathway,Pathway, Biosynthetic,Pathways, Biosynthetic
D059765 Homologous Recombination An exchange of DNA between matching or similar sequences. Homologous Recombinations,Recombination, Homologous,Recombinations, Homologous
D060847 Metabolic Engineering Methods and techniques used to genetically modify cells' biosynthetic product output and develop conditions for growing the cells as BIOREACTORS. Engineering, Metabolic
D019207 beta Carotene A carotenoid that is a precursor of VITAMIN A. Beta carotene is administered to reduce the severity of photosensitivity reactions in patients with erythropoietic protoporphyria (PORPHYRIA, ERYTHROPOIETIC). Betacarotene,BellaCarotin,Carotaben,Max-Caro,Provatene,Solatene,Vetoron,beta-Carotene,Carotene, beta,Max Caro,MaxCaro
D025062 Yarrowia A genus of ascomycetous yeast in the family Dipodascaceae, order SACCHAROMYCETALES. Yarrowia lipolytica,Yarrowias

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