Molecular cloning and characterization of a cDNA encoding a larval cuticle protein of Bombyx mori.
ABSTRACT Cuticle proteins termed LCPs are the major protein components of the larval integument of the silkworm, Bombyx mori. We purified an 18 kDa LCP (LCP18) from the guanidine hydrochloride extract of the larval cuticle and identified an LCP18 cDNA clone. The deduced primary structure and mRNA expression pattern of LCP18 are similar to those of other Bombyx LCPs and to several cuticle proteins identified in other insect species. RNA blot analysis demonstrated that the biosynthesis of LCP18 is regulated in a stage-dependent manner at the level of mRNA in epidermal cells. An in vivo study using a juvenile hormone analogue suggested that juvenile hormone positively regulates expression of LCP18 mRNA during larval intermolt stages.
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ABSTRACT: A cuticle protein gene, PbLCP12.1, from the white spotted flower chafer, Protaetia brevitarsis, was isolated and characterized. The gene contains an ORF of 336 nucleotides capable of encoding a 113 amino acid polypeptide with a predicted molecular mass of 12,138 Da and pI of 4.15. The PbLCP12.1 protein contained a type-specific consensus sequence identifiable in other insect cuticle proteins. The deduced amino acid sequence of the PbLCP12.1 cDNA is most similar to Bombyx mori cuticle protein BmLCP18 (37 protein sequence identity). Northern blot analysis revealed that PbLCP12.1 showed the epidermis-specific expression.01/2005; 11(1).
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ABSTRACT: In a previous study, three larval cuticle protein genes were cloned from the mulberry longicorn beetle, Apriona germari (Comp. Biochem. Physiol. B 136, 803-811, 2003). In the present study, the genomic structures of these three larval cuticle protein genes (AgLCP9.2, AgLCP12.6 and AgLCP12.3) were elucidated. All three cuticle protein genes consist of one intron and two exons. Southern blot analysis of genomic DNA suggested that three cuticle protein genes are a single copy gene. In addition, a novel larval cuticle protein gene, AgLCP10.6, was cloned from A. germari in this study. The AgLCP10.6 cDNA contains an ORF of 300 nucleotides that are capable of encoding a 100-amino acid polypeptide with a predicted molecular mass of 10.6 kDa. The amino acid sequence deduced from the AgLCP10.6 cDNA contained a type-specific consensus sequence identifiable in other insect cuticle proteins and is most homologous to Drosophila melanogaster cuticle protein ACP65A (51 % protein sequence identity). Northern blot analysis revealed that AgLCP10.6 showed epidermis-specific expression.01/2007; 14(1).
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ABSTRACT: In our research to identify gene involved in the cuticle protein, we cloned a novel cuticle protein gene, ApCP15.5, from the Chinese oak silkmoth, Antheraea pernyi, larvae cDNA library. The gene encodes a 149 amino acid polypeptide with a predicted molecular mass of 15.5 kDa and a pI of 9.54. The ApCP15.5 contained a type-specific consensus sequence identifiable in other insect cuticle proteins and the deduced amino acid sequence of the ApCP15.5 cDNA is most homologous to Tenebrio molitor-C1B ( protein sequence identity), followed by Locusta migratoria-76 ( protein sequence identity). Northern blot and Western blot analyses revealed that the ApCP15.5 showed the epidermis-specific expression. The expression profile of ApCP15.5 indicated that the ApCP15.5 mRNA expression was detected in the early stages after larval ecdysis and larval-pupal metamorphosis, and its expression level was most significant on the first day of larval ecdysis and pupal stage. The ApCP15.5 was expressed as a 15.5 kDa polypeptide in baculovirus-infected insect cells.01/2005; 10(1).