Molecular cloning of theARG7gene ofSchizosaccharomyces pombeencoding argininosuccinate lyase

✍ Scribed by Jacques E. Remacle; Didier Breyer; Roland Loppes

• Publisher: Springer-Verlag
• Year: 1988
• Tongue: English
• Weight: 410 KB
• Volume: 14
• Category: Article
• ISSN: 0172-8083
• DOI: 10.1007/bf00419996

✦ Synopsis

A gene bank of Sau3A partially restricted Schizosaccharomyces pombe DNA in YEpl3 was used to transform an arg4 mutant ofSaccharomyces cerevisiae. One colony was recovered which contained the YEpl3 plasmid bearing a large insert complementing the argininosuccinate lyase (ASL) mutation. As shown by restriction mapping and subcloning experiments, the DNA sequence required for complementation is localized on a 2 kb BamHI-BamHI fragment. The plasmid complemented several S. cerevisiae arg4 mutants of independent origin and a S. pombe arg7 mutant lacking ASL. Low but significant ASL activities were detected in crude extracts of these transformants. No complementation of the E. coli argH mutant was observed. Southern blot hybridizations showed that the insert originates from the S. pombe genome. No cross-hybridization was found between this sequence and S. cerevisiae DNA. It can be concluded that the cloned DNA fragment bears the S. pombe ARG7 gene coding for ASL.