Screening of cDNA libraries from grandular trichomes of Salvia fruticosa and Salvia pomifera by colony - low stringency hybridization to identify terpene synthase genes
Owsiak, S.
Both cDNAs libraries of Salvia fruticosa and Salvia pomifera previously created as a part of the INTESY project, were screened in order to identify new terpene synthase candidates. The colony hybridization technique was employed utilizing three radioactive probes of monoterpene synthases, probes made from cineol, sabinen and 215 cDNAs, which had been identified in previous studies. The colony hybridization resulted in a total number of 52 cDNA clones chosen for further investigation. After sequencing and comparison of the translated query against protein database available on a public domain, the two cDNA clones (hybridized with 215 radioactive probe) showed high similarity with terpene synthases, although both of them were only partial. Bioinformatic work revealed that the two isolated clones were a part of the same gene, 3-carene synthase homologue. The longest open reading frame encoded a deduced amino acid sequence, about 220 residues. The longest open reading frame encoded a deduced amino acid sequence, about 220 residues. Multiple protein alignment with other monoterpene synthases showed that the deduced polypeptide fragment was a part of the active site domain.
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