title: Expression of cellulase genes in Saccharomyces cerevisiae via δ-integration subject to auxotrophic markers.
abstract: δ-Integration can improve the expression stability and increase the copy number of exogenous genes in Saccharomyces cerevisiae. Generally, δ-integration vectors employ auxotrophic markers, such as LEU2-d, HIS3, TRP1, and URA3, to screen transformants. In this study, two cellulase genes (β-glucosidase and endoglucanase) were integrated into the S. cerevisiae W303-1A chromosome as reporters, by δ-integration with the aforementioned auxotrophic markers. Eight strains, L-BGL, H-BGL, T-BGL, U-BGL, L-EG, H-EG, T-EG, and U-EG, were selected and tested. After 5 days growth, cellulase activities (U ml(-1)) were 3, 2.3, 1.8, 1.5, 29, 12, 8, and 1.5, respectively. The results showed that auxotrophic markers influenced the expression of cellulase genes.
authors.affiliation: Tianjin R&D Center for Petrochemical Technology, Tianjin University, Tianjin, 300072, China. yi1mi2@163.com
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.forename: Cheng
authors.initials: C
authors.lastname: Liu
authors.forename: Shaolan
authors.initials: S
authors.lastname: Zou
authors.forename: Huajun
authors.initials: H
authors.lastname: Yang
authors.forename: Jiefang
authors.initials: J
authors.lastname: Hong
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ma
authors.forename: Minhua
authors.initials: M
authors.lastname: Zhang
mesh_terms: D044602:Cellulases / Q000096:biosynthesis* / Q000235:genetics*; D018628:Gene Dosage; D015870:Gene Expression; D005819:Genetic Markers; D060847:Metabolic Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics*; D012641:Selection, Genetic; D014170:Transformation, Genetic
chemical_list: D005819:Genetic Markers; D011994:Recombinant Proteins; D044602:Cellulases