Isolation and identification of microsatellite repeat motifs from the Epinephelus fuscoguttatus genome

Epinephelus fuscoguttatus belongs to one of the largest serranidae fish family. Genetic information regarding existing fish populations in the wild is crucial for the conservation, particularly since the species is listed under the IUCD Red List due to intense fishing. Microsatellites of E. fuscoguttatus were isolated using streptavidin-biotin enrichment method. In total, 378 microsatellites were identified and characterized. Of these 378 total microsatellites, 46 (12.2%) were mononucleotides, 175 (46.3%) were dinucleotides, 109 (28.8%) were trinucleotides, 36 (9.5%) were tetranucloetides, 7 (1.9%) were pentanucleotides, 4 (1.1%) were hexanucleotides and 1 (0.3%) was a heptanucleotide. The most abundant microsatellite present in E. fuscoguttatus was the dinucleotide motif, (AC)n.Key words: Enrichment, Epinephelus fuscoguttatus, microsatellite, diversity

Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant

N-glycans are biologically important oligosaccharides associated with the asparagine residue that may exist in protein-bound or unbound forms in all eukaryotes (including yeasts) and some bacteria. The- core structure of these oligosaccharides is based on the trimannosyl chitobiose structure resulting from cellular N-glycosylation. Preparative-scale amounts of these oligosaccharides are important for chemical, structural and functional studies due to their biological significance. Therefore, we explored a biochemical approach of oligosaccharide preparation using mutant-derived monoglucosylated lipid-linked oligosaccharides (LLOs) required for the assembly of N-linked glycoproteins and non-monoglucosylated free-oligosaccharides (fOSs) from misfolded N-linked glycoproteins using an N-glycosylation (alg) mutant of Saccharomyces cerevisiae. Oligosaccharide extracts of fOSs and LLOs from the alg8 S. cerevisiae mutant lacking the ALG8 gene were profiled using fluorescence- and evaporative light scattering-based HPLC. LLOs did not produce accumulated levels of the target mutant- related monoglucosylated (Glc1Man9GlcNAc2) at 100 ml scale. However, it was possible to detect truncated oligomannose (paucimannose) structures in the fOSs of the alg8 mutant

Fluorescence and evaporative light scattering HPLC profiling of intracellular asparagine (N)-linked oligosaccharides from Saccharomyces cerevisiae using the alg8 mutant

N-glycans are biologically important oligosaccharides associated with the asparagine residue that may exist in protein-bound or unbound forms in all eukaryotes (including yeasts) and some bacteria. The- core structure of these oligosaccharides is based on the trimannosyl chitobiose structure resulting from cellular N-glycosylation. Preparative-scale amounts of these oligosaccharides are important for chemical, structural and functional studies due to their biological significance. Therefore, we explored a biochemical approach of oligosaccharide preparation using mutant-derived monoglucosylated lipid-linked oligosaccharides (LLOs) required for the assembly of N-linked glycoproteins and non-monoglucosylated free-oligosaccharides (fOSs) from misfolded N-linked glycoproteins using an N-glycosylation (alg) mutant of Saccharomyces cerevisiae. Oligosaccharide extracts of fOSs and LLOs from the alg8 S. cerevisiae mutant lacking the ALG8 gene were profiled using fluorescence- and evaporative light scattering-based HPLC. LLOs did not produce accumulated levels of the target mutant- related monoglucosylated (Glc1Man9GlcNAc2) at 100 ml scale. However, it was possible to detect truncated oligomannose (paucimannose) structures in the fOSs of the alg8 mutant