Identification, isolation and bioinformatic analysis of squalene sinthase-like cDNA fragments in „Botryococcus terribilis” AICB 870 strain
Keywords:
Bioinformatics analysis, Botryococcus terribilis, cDNA, hydrocarbons, squalene synthase-like.Abstract
Botryococcus terribilis is a freshwater colonial green microalga very similar to B. braunii, due to the hydrocarbon biosynthesis and accumulation of those biosynthetic products in the extracellular matrix. The hydrocarbon biosynthesis pathway was intensively studied, especially in differents strains of B. braunii. Recent studies revealed the presence of three squalene synthase-like (SSL) enzymes involved in the last steps of hydrocarbon biosynthesis from B. braunii. The aim of the study is to identify homologous SSL enzymes to B. terribilis AICB 870, a freshwater isolate from Bihor County (Romania), based on new isolated cDNA fragments and bioinformatics analysis of sequenced fragments. Light and fluorescence microscopy observation revealed that AICB 870 strain presents features similar to a B. terribilis species, especially simple or branched mucilaginous processes and a high number of lipids vesicles. PCR primers designed using SSL nucleotide sequences from B. braunii were successfully used to amplify homologous SSL cDNA fragment in the AICB 870 strain. Bioinformatic analysis of nucleotides and translated amino acid sequences including G+C content, nucleotide frequencies, amino acids frequencies, computed Mw/pI and transmembrane motif prediction showed a high degree of similarity between the SSL identified as pertaining to Botryococcus braunii and those generated in the present work. The results of the present study pointed out for the first time the presence of three squalene synthase-like enzymes in a strain of B. terribilis species.
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