To elucidate fungal diversity in methane hydrate-bearing deep-sea marine sediments in the South China Sea, internal transcribed spacer (ITS) regions of rRNA genes from five different sediment DNA samples were amplified and phylogenetically analyzed. Total five ITS libraries were constructed and 413 clones selected randomly were grouped into 24 restriction patterns by Amplified Ribosomal DNA Restriction Analysis (ARDRA). ITS sequences of 44 representative clones were determined and compared with the GenBank database using gapped-BLAST. The phylogenetic analysis showed that the ITS sequences (71-97% similarity) were similar to those of Phoma, Lodderomyces, Malassezia, Cryptococcus, Cylindrocarpon, Hortaea, Pichia, Aspergillus and Candida. The remaining sequences were not associated to any known fungi or fungal sequences in the public database. The results suggested that methane hydrate-bearing deep-sea marine sediments harbor diverse fungi. This is the first report on fungal communities from methane hydrate-bearing deep-sea marine sediments in South China Sea.
The South China Sea, which is one of the largest marginal seas in the world, is predicted to have suitable accumulation conditions and exporting prospects for natural gas hydrate. The aim of this study was to explore the bacterial community composition of deep-sea sediments from such an ecosystem. DNA was extracted by five different methods and used as templates for PCR amplification of the V3 regions of the 16S rRNA gene. Denaturing gradient gel electrophoresis (DGGE) was used to separate the amplified products and analyse the 16S rRNA gene diversity of sediment samples. The results of DGGE indicated that the bacterial community composition is influenced by DNA extraction methods. Sequencing dominant bands demonstrated that the major phylogenetic groups identified by DGGE belong to Proteobacteria, Bacteroidetes, gram-positive bacteria and Archaea. Integrating different DNA extraction procedures are needed to analyse the actual bacterial diversity from environment when the amplification of 16S rRNA gene and construction of representative clone library were adopted. AbbreviationsDGGE denaturing gradient gel electrophoresis mbsf metres below the seafloor EL Enzymatic lysis method CL Chemical lysis method SN 3S DNA isolation Kit for environmental samples of Shenergy biocolor Company IM improved DNA extraction method
Edible bird's nest (EBN) is made of the swiftlets' saliva, which has attracted rather more attention owing to its nutritious and medical properties. Although protein constitutes the main composition and plays an important role in EBN, few studies have focused on the proteomic profile of EBN. The purpose of this study was to produce a proteomic map and clarify common EBN proteins. Liquid-phase isoelectric focusing (LIEF) was combined with two-dimensional electrophoresis (2-DE) for comprehensive analysis of EBN proteins. From 20 to 100 protein spots were detected on 2-DE maps of EBN samples from 15 different sources. The proteins were mainly distributed in four taxa (A, B, C, and D) according to their molecular mass. Taxa A and D both contained common proteins and proteins that may be considered another characteristic of EBN. Taxon A was identified using MALDI-TOF-TOF/MS and found to be homologous to acidic mammalian chitinase-like ( Meleagris gallopavo ), which is in glycosyl hydrolase family 18.
The proliferation of fake and inferior edible bird's nest (EBN) products has recently become an increasingly serious concern. To identify and classify EBN products, a competitive enzyme-linked immunoassay (ELISA) was developed to quantitate sialoglycoprotein in EBN used in food and cosmetic applications. The characteristic sialoglycoprotein in EBN was found, extracted, purified, and analyzed. Sialoglycoprotein, considered the main carrier of sialic acid in EBN, consisted of 106 and 128 kDa proteins. A monoclonal antibody that could recognize both proteins was prepared. The heat-treated process did not change the affinity of sialoglycoprotein with the antibody. An optimized ELISA method was established with a cross-reactivity of less than 0.1% and an IC(50) of 3.3 μg/mL. On the basis of different food and cosmetic samples, the limits of detection (LOD) were 10-18 μg/g. Recoveries of fortified samples at levels of 20 and 80 μg/g ranged from 81.5 to 96.5%, respectively. The coefficients of variation were less than 8.0%.
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