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Identification and expression analysis of minichromosome maintenance proteins in the silkworm, Bombyx mori.

ABSTRACT: The minichromosome maintenance protein (MCM) family is involved in the regulatory role of DNA replication in eukaryotic organisms. A cDNA encoding of an MCM of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced. The resultant amino acid sequence and phylogenetic analysis revealed high identity to MCM, and specifically to MCM7, of vertebrates and invertebrates. An RT-PCR showed that the bmMCM7 transcript was present in the ovaries, testes, silk glands, and fat bodies of larval silkworms. Expression plasmids were transformed into competent Escherichia coli and overexpressed. This is the first report on the identification of MCM helicase of the silkworm, B. mori.

SUBMITTER: Yamamoto K

PROVIDER: S-EPMC3016899 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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Identification and expression analysis of minichromosome maintenance proteins in the silkworm, Bombyx mori.

Yamamoto Kohji K Aso Yoichi Y Fujii Hiroshi H

Journal of insect science (Online) 20100101

The minichromosome maintenance protein (MCM) family is involved in the regulatory role of DNA replication in eukaryotic organisms. A cDNA encoding of an MCM of the silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequenced. The resultant amino acid sequence and phylogenetic analysis revealed high identity to MCM, and specifically to MCM7, of vertebrates and invertebrates. An RT-PCR showed that the bmMCM7 transcript was ...[more]

PMID: 21070176

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Project description:BACKGROUND: Carboxylesterase is a multifunctional superfamily and ubiquitous in all living organisms, including animals, plants, insects, and microbes. It plays important roles in xenobiotic detoxification, and pheromone degradation, neurogenesis and regulating development. Previous studies mainly used Dipteran Drosophila and mosquitoes as model organisms to investigate the roles of the insect COEs in insecticide resistance. However, genome-wide characterization of COEs in phytophagous insects and comparative analysis remain to be performed. RESULTS: Based on the newly assembled genome sequence, 76 putative COEs were identified in Bombyx mori. Relative to other Dipteran and Hymenopteran insects, alpha-esterases were significantly expanded in the silkworm. Genomics analysis suggested that BmCOEs showed chromosome preferable distribution and 55% of which were tandem arranged. Sixty-one BmCOEs were transcribed based on cDNA/ESTs and microarray data. Generally, most of the COEs showed tissue specific expressions and expression level between male and female did not display obvious differences. Three main patterns could be classified, i.e. midgut-, head and integument-, and silk gland-specific expressions. Midgut is the first barrier of xenobiotics peroral toxicity, in which COEs may be involved in eliminating secondary metabolites of mulberry leaves and contaminants of insecticides in diet. For head and integument-class, most of the members were homologous to odorant-degrading enzyme (ODE) and antennal esterase. RT-PCR verified that the ODE-like esterases were also highly expressed in larvae antenna and maxilla, and thus they may play important roles in degradation of plant volatiles or other xenobiotics. CONCLUSION: B. mori has the largest number of insect COE genes characterized to date. Comparative genomic analysis suggested that the gene expansion mainly occurred in silkworm alpha-esterases. Expression evidence indicated that the expanded genes were specifically expressed in midgut, integument and head, implying that these genes may have important roles in detoxifying secondary metabolites of mulberry leaves, contaminants in diet, and odorants. Our results provide some new insights into functions and evolutionary characteristics of COEs in phytophagous insects.

Dataset Information

Identification and expression analysis of minichromosome maintenance proteins in the silkworm, Bombyx mori.

Publications

Identification and expression analysis of minichromosome maintenance proteins in the silkworm, Bombyx mori.

Similar Datasets

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