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Characterization of Expressed Sequence Tags (ESTs) Generated from the Bombyx mandarina Whole Larvae and Molecular Cloning of Serine Protease Homologue Gene  

Hwang, Jae Sam (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Yun, Eun Young (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Goo, Tae Won (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Kim, Iksoo (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Choi, Kwang Ho (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Seong, Su Il (Department of Life Science, Collage of Natural Science, The University of Suwon)
Kim, Keun Young (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Lee, Sang Mong (Department of Sericultural and Entomology, Faculty of Agriculture, Miryang National University)
Kang, Seok Woo (Department of Agricultural Biology, National Institute of Agricultural Science and Technology)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.9, no.2, 2004 , pp. 167-171 More about this Journal
Abstract
We constructed an oligo-d(T) primed directional cDNA library from the Bombyx mandarina whole larvae. In an effort to isolate genes expressed in the B. mandarina, 227 expressed sequence tags (ESTs) were generated by single-pass sequencing from the cDNA library. Sequence analysis showed that 107 clones (47.1%) were classified into known genes and 120 clones (52.9%) were novel transcripts, which are unknown for their function. Of the 107 known genes, the most abundant gene was found to be actin and followed by serine protease in the expression profile. Among these clones, a serine protease homolog (BmSP) which is a class of proteolytic enzymes isolated. Full-length sequence of the BmSP cDNA clone was 922 bp in length and has an open reading frame of 276 amino acids. The conserved histidine, aspatic acid and serine residues forming the catalytic center as well as cysteine residues contributing to three disulphide bonds also were found in Bmsp gene. mRNA expression analysis revealed a high and specific expression of the gene only in midgut tissue, suggesting that BmSP gene is closely associated with the expression of digestive enzyme.
Keywords
ESTs; Bombyx mandarina; Serine protease; Midgut;
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