Agarum clathratum, brown algae, play important ecological roles in marine ecosystem, but can cause secondary environ-ment pollution when they pile up on the beach. In order to resolve the environment problem by A. clathratum, we focus to isolate and identify Penicillium because many species are well known to produce extracellular enzymes. A total of 32 Penicillium strains were isolated from A. clathratum sam-ples that collected from 13 sites along the mid-east coast of Korea in summer. They were identified based on morpho-logical characters and phylogenetic analysis using β-tubulin DNA sequences as well as a combined dataset of β-tubulin and calmodulin. A total of 32 strains were isolated and they were identified to 13 Penicillium species. The commonly iso-lated species were Penicillium citrinum, P. roseomaculatum, and Penicillium sp. Among 13 Penicillium species, four spe-cies – P. bilaiae, P. cremeogriseum, P. madriti, and P. rose-omaculatum – have not been previously recorded in Korea. For these four new species records to Korea, we provide mor-phological characteristics of each strain.
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Five cDNAs encoding two α-tubulins(α13 and α15), two β-tubulins(β5 and β5), and one flagellar calmodulin (Cal-1) were cloned from naegleria gruberi NB-1 and their nt sequences were determined. The α13(EMBL number X81049) and β1(EMBL number X81050) contained a complete open reading frame for α-tubulin and β-tubulin, respectively. The other three clones (α15, β5 and Cal-1) had a part of coding region and a 3’ untranslated region of the respective genes. The α13, β1 and Cal-1 had no homologous sequences in the coding regions and in the 3’ untranslated sequences. However, the α13 and β1 shared an eight nucleotide (AATACAAA) sequence in front of the respective initiation codons. The AATACAAA sequence was also found in N. gruberi strain NEG α-tubulin cDNA clone(αpT1) at the same position. Comparison of the α13 to the αpT1 revealed another stretch of identical sequence, which is 30 nts long, in the 5’ untranlated region.
Two novel calcium-binding proteins, named CAB-I and CAB-II, have been isolated from Streptomyces coelicolor. Purification of the calcium-binding proteins involved heat treatment, fractionation with ammonium sulfate, acid treatment, anion exchange and hydrophobic interaction column chromatography, FPLC gel filtration, and preparative isoelectric focusing. A chelex competitive assay and ^45Ca autoradiography verified the calcium-binding ability of the proteins. The major band CAB-II has an apparent molecular weight of 26,000 determined by SDS-polyacrylamide gel electrophoresis and 340,000 determined by gel filtration. The isoelectric point of this molecule showed the acidic nature of the molecule. N-terminal amino acid sequence analysis shows homology to rat Ca^2+/calmodulin-dependent protein kinase-II (CAB-II) and yeast phosphoprotein phosphatase (CAB-I).