Wyniki wyszukiwania

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A simple and reliable method for DNA extraction from bivalve mantle

100%

Aranishi F., Okimoto T.

Journal of Applied Genetics

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2006

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tom 47

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nr 3

A simple and reliable method was developed for extracting genomic DNA from preserved mantle tissues of Pacific oyster Crassostrea gigas for reproducible PCR amplification. The method was based on destruction of the tissue using Proteinase K, Chelex 100 resin, detergents, and urea, followed by preferential capturing of genomic DNA with silica particles. Approximately 5 mg of mantle tissue provided a sufficient quality and quantity of DNA for several hundreds of PCR reactions amplifying the hypervariable mitochondrial DNA intergenic spacer, which is a useful genetic marker for population structure analysis of Pacific oyster. The method can be applied for DNA preparation from not only fresh and frozen but also ethanol-preserved mantle tissues, so this rapid and economical method can serve for investigating a large number of bivalve specimens collected in the field and next transported in ethanol at ambient temperature.

2

Sequence polymorphism in a novel noncoding region of Pacific oyster mitochondrial DNA

100%

Aranishi F., Okimoto T.

Journal of Applied Genetics

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2005

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tom 46

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nr 2

Nucleotide sequence polymorphism in a 641-bp novel major noncoding region of mitochondrial DNA (mtDNA-NC) of the Pacific oyster Crassostrea gigas was analysed for 29 cultured individuals within the Goseong population. A total of 30 variable sites were detected, and the relative frequency of nucleotide alteration was determined to be 4.68. Alterations were mostly single nucleotide substitutions. Transition, transversion, both transition and transversion, and both transversion and nucleotide deletion were observed at 18, 9,2 and 1 sites, respectively. Among 29 specimens, 22 haplotypes were identified, and pairwise genetic diversity of haplotypes was calculated to be 0.988 from multiple sequence substitutions using the two-parameter model. A phylogenetic tree, obtained for haplotypes by the neighbor-joining method, showed a single cluster of linkages. The cluster comprised 11 haplotypes associating with 14 specimens, while the other 11 haplotypes associating with 15 specimens were scattered. This mtDNA-NC presenting a high nucleotide sequence polymorphism is a potential mtDNA control region. It therefore can serve as a genetic marker for intraspecies phylogenetic analysis of the Pacific oyster and is more useful than the less polymorphic mtDNA coding genes.

3

Genetic relationship between cultured populations of Pacific oyster revealed by RAPD analysis

100%

Aranishi F., Okimoto T.

Journal of Applied Genetics

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2004

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tom 45

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nr 4

4

Identification of gadoid species [Pisces, Gadidae] by PCR-RFLP analysis

80%

Aranishi F., Okimoto T., Izumi S.

Journal of Applied Genetics

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2005

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tom 46

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nr 1

A rapid PCR-RFLP analysis was optimized to identify the presence of 3 closely related gadoid fish species: Alaska pollack Theragra chalcogramma, Pacific cod Gadus macrocephalus and Atlantic cod Gadus morhua in commercial seafood products. Gadoid universal primers were designed for PCR amplification of a 558-bp fragment encoding the mitochondrial cytochrome b gene. Without purification of the PCR products, double digestion with Eco32l and Eco1051 restriction enzymes generated reproducible species-specific restriction patterns visualizing 3 fragments (106 bp, 161 bp and 291 bp) in Alaska pollack and 2 fragments (106 bp and 452 bp) in Pacific cod, whereas no cleavage was observed in Atlantic cod. This PCR-RFLP analysis is simple, rapid and reliable, and therefore can be routinely applied to discover fraudulent substitution among 3 economically important gadoid species in commercial seafood products.

Ograniczanie wyników

4 Journal of Applied Genetics

4 Aranishi F.

4 Okimoto T.

1 Izumi S.

1 2006

2 2005

1 2004

A simple and reliable method for DNA extraction from bivalve mantle

Sequence polymorphism in a novel noncoding region of Pacific oyster mitochondrial DNA

Genetic relationship between cultured populations of Pacific oyster revealed by RAPD analysis

Identification of gadoid species [Pisces, Gadidae] by PCR-RFLP analysis