Characterization of Expressed Sequence Tags (ESTs) Generated from the Bombyx mandarina Whole Larvae and Molecular Cloning of Serine Protease Homologue Gene
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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 eading frame of 276 amino acids. The conserved histidine, aspatic acid and serine resihues forming the catalytic center as well as cysteine residues contributing to three disulphide bonds also were found in Bmsp ene. 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.
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