Cloning and sequence analysis ofarom gene fromSclerotinia sclerotiorum

Abstract

Anarom gene was cloned from genomic DNA ofScleortinia sclerotiorum by inverse PCR. The evolutionary relationships ofS. sclerotiorum and other fungi inarom gene were studied. Results showed that thearom gene from ofS. sclerotiorum has a single open reading frame of 4773 bp and does not include any introns. The derived amino acid sequence consists of 1 590 residues, and it is homologous to all fungal AROM proteins studied so far. The theoretical isoelectric point (pI) and molecular weight (Mw) is 6.5 and 172.66 kD, respectively. GC percentage of thearom gene is 44.94. According to the results of searching from CDD and Prosite database, AROM protein ofS. sclerotiorum contains five conserve domains: 3-dehydroquinate synthase domain, 3-dehydroquinate dehydratase (3-dehydroquinase) domain, shikimate 5-dehydrogenase domain, shikimate kinase domain, and-enolpyruvylshikimate-3-phosphate synthase (EPSP sythase) domain, and four motifs: two EPSP synthase signatures, dehydroquinase class I active site, shikimate kinase signature. According to the PIR Site Rule PIRSR000514-1, four functionally important amino acid residues are found by alignment. Putative TATA box and CAAT box locate separately in −23 and −77 loci in 5′ un-translated region, and two loci found in downstreamarom gene are likely polyadenylation signals. In addition, phylogeny ofarom gene is analyzed.

Keywords

Sclerotinia sclerotiorum / arom / Gene cloning / Q781 / A

References

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