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 re
Sequence analysis of theARG7gene ofSchizosaccharomyces pombecoding for argininosuccinate lyase
โ Scribed by Roland Loppes; Reiner Michels; Isabelle Decroupette; Bernard Joris
- Publisher
- Springer-Verlag
- Year
- 1991
- Tongue
- English
- Weight
- 825 KB
- Volume
- 19
- Category
- Article
- ISSN
- 0172-8083
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โฆ Synopsis
ARG7 gene, coding for argininosuccinate lyase (EC 4.3.2.1), in the fission yeast (Schizosaccharomyces pombe) has been determined. It consists of an open reading frame of 461 codons. The deduced protein has a molecular weight of 51 200 Da. The gene is devoid of introns which is confirmed by the fact that it is expressed in Escherichia coli after spontaneous insertion of a bacterial sequence probably bearing a prokaryotic promoter. A perfect "TATA" box is found at 72 and the major transcription initiation site in Saccharomyces cerevisiae is located at -11 as shown by primer extension experiments. Comparison of the S. pombe lyase with related proteins from other organisms reveals an important degree of conservation except in the carboxyterminal part of the polypeptide. Additionally, a deletion removing 66 amino acids of the carboxy terminus yields an enzyme exhibiting some biological activity. A unique 1 500 b transcript was found in S. cerevisiae when the intact gene was present, but the deleted version of the gene gave rise to at least three transcripts of 1 800, 2 800 and 3 900 b.
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