Cloning, sequencing and characterization of the alpha-aminoadipate reductase gene (LYS2) from Saccharomycopsis fibuligera.
The gene putatively encoding alpha-aminoadipate reductase (AAR) was isolated successfully by degenerate PCR and chromosome walking, based on cassette PCR methods, from the dimorphous yeast Saccharomycopsis fibuligera PD70 and was named SfLYS2. Sequence analysis revealed that it contained a putative open reading frame (ORF) of 4161 bp and encoded a polypeptide of 1386 amino acids. The deduced translation product shared an identity of 53% and 51% to the Lys2p homologues of Candida albicans and Saccharomyces cerevisiae, respectively. An atypical TATA box and a GCN4-box element were found in the 5'-upstream region. Genomic Southern hybridization suggested the presence of a single locus of SfLYS2 in the S. fibuligera genome. Expression of the ORF of SfLYS2 in a lys2(-) strain of S. cerevisiae could functionally complement the lysine mutant of the S. cerevisiae strain. S. fibuligera could use lysine as the sole nitrogen source but its growth was inhibited on the alpha-aminoadipate (AA) medium. Approximately 90% of the mutants of S. cerevisiae resistant to AA are lysine auxotrophs; in contrast all the mutants of S. fibuligera resistant to AA recovered in this work were not lysine auxotrophs.
Yan
Hua
H
Centre of Microbial Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, People's Republic of China.
He
Peng
P
Cheng
Hai-Rong
HR
Shen
An
A
Jiang
Ning
N
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D050647:L-Aminoadipate-Semialdehyde Dehydrogenase / Q000235:genetics*; D008969:Molecular Sequence Data; D009088:Mucoproteins; D016366:Open Reading Frames; D010802:Phylogeny; D012442:Saccharomycopsis / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D017386:Sequence Homology, Amino Acid
D003470:Culture Media; D005656:Fungal Proteins; D009088:Mucoproteins; C010393:lysin, gastropoda; D050647:L-Aminoadipate-Semialdehyde Dehydrogenase