Cloning and expression analysis of the Bombyx mori α-amylase gene (Amy) from the indigenous Thai silkworm strain, Nanglai
1The Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, Thailand
2The National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8634, Japan
3The Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, Thailand
4These two authors contributed equally to this work.
Abstract
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 81-54% identity to other insect α‐ 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.
Keywords: Amy, Bombyx mori, GAL4-UAS, silkworm
Abbreviations: Amy, α-amylase gene; BmAMY, Bombyx mori α-amylase enzyme
Correspondence:
a yingjung_@hotmail.com,
b isaorj@affrc.go.jp,
c fsciarp@ku.ac.th,
d fscistr@ku.ac.th,
e ttamura@affrc.go.jp,
f* khawlln@yahoo.com,*Corresponding author
Editor: Craig Coates was editor of this paper
Received: 23 February 2010 | Accepted: 5 June 2010 | Published: 29 March 2011
Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.
ISSN: 1536-2442 | Volume 11, Number 38
Ngernyuang N, Kobayashi I, Promboon A, Ratanapo S, Tamura T, Ngernsiri L. 2011. Cloning and expression analysis of the Bombyx mori α-amylase gene (Amy) from the indigenous Thai silkworm strain, Nanglai. Journal of Insect Science 11:38 available online: insectscience.org/11.38
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