Compagno C, Tura A, Ranzi B M, Alberghina L, Martegani E
Dipartimento di Fisiologia e Biochimica Generali, Universitá di Milano, Italy.
Biotechnol Prog. 1993 Nov-Dec;9(6):594-9. doi: 10.1021/bp00024a005.
Efficient expression of a foreign gene requires a stable vector present at a high number of copies per cell. We have constructed an autoselection system for the stable maintenance of expression vector in the yeast Saccharomyces cerevisiae that uses the fructose 1,6-bisphosphate aldolase gene (FBA1) to stabilize plasmids in cells bearing a disruption of the chromosomal FBA1 gene. This system allowed us to obtain stable production of a reporter heterologous enzyme (Escherichia coli beta-galactosidase) in rich media. By using an inducible promoter to regulate the expression of FBA1 gene, we have also obtained the modulation of plasmid copy number by carbon source.
外源基因的高效表达需要每个细胞中存在高拷贝数的稳定载体。我们构建了一种自动选择系统,用于在酿酒酵母中稳定维持表达载体,该系统利用1,6-二磷酸果糖醛缩酶基因(FBA1)来稳定携带染色体FBA1基因缺失的细胞中的质粒。该系统使我们能够在丰富培养基中稳定生产报告异源酶(大肠杆菌β-半乳糖苷酶)。通过使用诱导型启动子来调节FBA1基因的表达,我们还通过碳源实现了质粒拷贝数的调控。