RESUMO
The DREP-1 gene plays an important role in the process of apoptosis in Drosophila. The amino acid sequence of Drosophila ICAD was searched against the silkworm EST database. The resulted ESTs were clustered and assembled into a consensus sequence. Primers based on the consensus sequence were designed and the full length cDNA of ICAD gene, designated BmICAD, was cloned firstly from Bombyx mori. This BmICAD cDNA was 844-bp in length, containing a 522-bp open reading frame (ORF), which encoded a protein of 174 amino acids; the molecular weight of this protein was estimated to be 19.6 kDa and the isoelectric point was 4.23. The BmICAD protein shares 36% identity with Drosophila DREP-1. No signal has inspected by Northern blot, but RT-PCR analysis showed that BmICAD was testis-specific expressed at the mRNA level. Structure and clustering analysis indicated that this gene might be involved in the process of apoptosis that regulate the degradation of DNA.
Assuntos
Proteínas Reguladoras de Apoptose/genética , Bombyx/citologia , Clonagem Molecular , Expressão Gênica , Proteínas de Insetos/genética , Sequência de Aminoácidos , Animais , Apoptose , Proteínas Reguladoras de Apoptose/química , Proteínas Reguladoras de Apoptose/metabolismo , Sequência de Bases , Bombyx/classificação , Bombyx/genética , Bombyx/metabolismo , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Dados de Sequência Molecular , FilogeniaRESUMO
piggyBac is a short inverted terminal repeat (ITR) transposable element originally discovered in Trichoplusia ni. It is currently the preferred vector of choice for enhancer trapping, gene discovery and identifying gene function in insects and mammals. Many piggyBac-like sequences have been found in the genomes of phylogenetically species from fungi to mammals. We have identified 98 piggyBac-like sequences (BmPBLE1-98) from the genome data of domesticated silkworm (Bombyx mori) and 17 fragments from expressed sequence tags (ESTs). Most of the BmPBLE1-98 probably exist as fossils. A total of 21 BmPBLEs are flanked by ITRs and TTAA host dinucleotides, of which 5 contain a single ORF, implying that they may still be active. Interestingly, 16 BmPBLEs have CAC/GTG not CCC/GGG as the characteristic residues of ITRs, which is a surprising phenomenon first observed in the piggyBac families. Phylogenetic analysis indicates that many BmPBLEs have a close relation to mammals, especially to Homo sapiens, only a few being grouped with the T. ni piggyBac element. In addition, horizontal transfer was probably involved in the evolution of the piggyBac-like elements between B. mori and Daphnia pulicaria. The analysis of the BmPBLEs will contribute to our understanding of the characteristic of the piggyBac family and application of piggyBac in a wide range of insect species.
Assuntos
Bombyx/genética , Elementos de DNA Transponíveis/genética , Genoma de Inseto , Sequências Repetidas Terminais , Sequência de Aminoácidos , Animais , Evolução Molecular , Etiquetas de Sequências Expressas , Dados de Sequência Molecular , Filogenia , Homologia de Sequência de AminoácidosRESUMO
The fat body plays key roles in metabolism and sustenance of growth throughout the life of a silkworm, and thus represents a model tissue for studying development and metamorphosis. Analysis of 18,480 ESTs derived from larval and pupal fat body cDNA libraries allowed characterization of gene expression patterns in the fat body during metamorphosis. By Phrap assembling, 6,814 estimated transcription units (TUs) were generated, 2,673 of which contained 60% of the total ESTs and represented 1,366 distinctive known genes. These genes were classified into 14 categories based on their known or putative functions, and most were found to be involved in metabolism and cell defense. Further comparative analysis of the expression profiles revealed that the gene expression patterns in the larval fat body strikingly differed from those in the pupal fat body. From this, we identified a subset of genes potentially associated with metamorphic events such as the formation or destruction of specific tissues, and simultaneously confirmed the stage-specific expression patterns of several identified genes using RT-PCR examination. This work will provide a valuable resource for studying regulatory mechanisms associated with fat body developmental changes and silkworm metamorphosis.
Assuntos
Bombyx/crescimento & desenvolvimento , Bombyx/genética , Corpo Adiposo/metabolismo , Perfilação da Expressão Gênica , Metamorfose Biológica/genética , Animais , Primers do DNA/genética , Etiquetas de Sequências Expressas , Feminino , Genes de Insetos , Larva/genética , Masculino , Dados de Sequência Molecular , Pupa/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Análise de Sequência de DNARESUMO
According to the analysis of large scale EST sequencing of silkworm, Bombyx mori, we found that chi, glue and fruA of silkworm have very high homology at amino acid level and closely phylogenetic relative with that of microorganism, but lower similarity with genes of eelworm (Caenorhabditis elegans), fruitfly (Drosophila melanogaster), mosquito (Anopheles gambiae) and other relative insects, respectively. It indicates that each of them is likely to have common ancestor with that of microorganism. Namely, microbial genes were likely transferred to silkworm by horizontal gene transfer, instead of the vertical inheritance in evolutionary manner.
