Cloning and Expression Analysis of theBombyx mori α-amylaseGene (Amy) from the Indigenous Thai Silkworm Strain, Nanglai
Author(s) -
Nipaporn Ngernyuang,
Isao Kobayashi,
Amornrat Promboon,
Sunanta Ratanapo,
Toshiki Tamura,
Lertluk Ngernsiri
Publication year - 2011
Publication title -
journal of insect science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.551
H-Index - 49
ISSN - 1536-2442
DOI - 10.1673/031.011.0138
Subject(s) - biology , bombyx mori , bombycidae , open reading frame , foregut , complementary dna , bombyx , microbiology and biotechnology , gene , amylase , southern blot , sequence analysis , genetics , peptide sequence , biochemistry , enzyme , anatomy
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.
Accelerating Research
Robert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom
Address
John Eccles HouseRobert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom