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Cloning and expression analysis of the Bombyx mori ?-amylase gene (Amy) from the indigenous Thai silkworm strain, Nanglai.


ABSTRACT: ?-Amylase is a common enzyme for hydrolyzing starch. In the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), ?-amylase is found in both digestive fluid and hemolymph. Here, the complete genomic sequence of the Amy gene encoding ?-amylase from a local Thai silkworm, the Nanglai strain, was obtained. This gene was 7981 bp long with 9 exons. The full length Amy cDNA sequence was 1749 bp containing a 1503 bp open reading frame. The ORF encoded 500 amino acid residues. The deduced protein showed 81-54% identity to other insect ?-amylases and more than 50% identity to mammalian enzymes. Southern blot analysis revealed that in the Nanglai strain Amy is a single-copy gene. RT- PCR showed that Amy was transcribed only in the foregut. Transgenic B. mori also showed that the Amy promoter activates expression of the transgene only in the foregut.

SUBMITTER: Ngernyuang N 

PROVIDER: S-EPMC3281462 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature

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Cloning and expression analysis of the Bombyx mori α-amylase gene (Amy) from the indigenous Thai silkworm strain, Nanglai.

Ngernyuang Nipaporn N   Kobayashi Isao I   Promboon Amornrat A   Ratanapo Sunanta S   Tamura Toshiki T   Ngernsiri Lertluk L  

Journal of insect science (Online) 20110101


α-Amylase is a common enzyme for hydrolyzing starch. In the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), α-amylase is found in both digestive fluid and hemolymph. Here, the complete genomic sequence of the Amy gene encoding α-amylase from a local Thai silkworm, the Nanglai strain, was obtained. This gene was 7981 bp long with 9 exons. The full length Amy cDNA sequence was 1749 bp containing a 1503 bp open reading frame. The ORF encoded 500 amino acid residues. The deduced protein showed 8  ...[more]

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