Uccelletti D, Farina F, Morlupi A, Palleschi C
Foundation Institut Pasteur-Fondazione Cenci-Bolognetti, Department of Developmental and Cell Biology, University of Rome La Sapienza, Italy.
Res Microbiol. 1999 Jan-Feb;150(1):5-12. doi: 10.1016/s0923-2508(99)80041-2.
We isolated spontaneous mutants resistant to sodium orthovanadate in the biotechnologically significant yeast Kluyveromyces lactis. Resistance behaved as a recessive character in all mutants analyzed. Four genes were defined by complementation analysis, from vga1 to vga4. These mutants showed defects in N-linked as well as O-linked glycosylation processes. In addition, the mutants exhibited sensitivity to the aminoglycoside hygromycin B and to calcofluor white, with the exception of vga4; this mutant grew in the presence of the antibiotic as well as the parental wild type and was resistant to calcofluor. The mutations were accompanied by alterations in the cell wall structure, as revealed by the delocalization of chitin, changes in cell shape and size and by the clumpy aspect of the cultures. The mutants isolated provide basic tools for molecular and cellular analysis of glycosylation processes in K. lactis.
我们在具有生物技术重要性的乳酸克鲁维酵母中分离出了对原钒酸钠具有抗性的自发突变体。在所有分析的突变体中，抗性表现为隐性性状。通过互补分析确定了四个基因，从vga1到vga4。这些突变体在N-连接和O-连接糖基化过程中表现出缺陷。此外，除vga4外，这些突变体对氨基糖苷类潮霉素B和荧光增白剂呈敏感性；该突变体在抗生素存在的情况下能够生长，与亲本野生型一样，并且对荧光增白剂具有抗性。突变伴随着细胞壁结构的改变，这通过几丁质的重新定位、细胞形状和大小的变化以及培养物的块状外观得以揭示。分离出的突变体为乳酸克鲁维酵母糖基化过程的分子和细胞分析提供了基础工具。
