title: Expression of genes derived from the genomic DNA fragments of the brown-winged green bug (Plautia stali) symbiont in Escherichia coli.
abstract: Many insect species harbour symbiotic microorganisms (symbionts) that are generally unculturable in media. To utilize symbionts as genome resources, we examined whether insect symbiont genes can be expressed in Escherichia coli. 144 plasmid clones were isolated from gene libraries, which were constructed from the genomic DNA of the intestinal bacterial symbiont in the brown-winged green bug, Plautia stali, using an E. coli system. Proteins prepared from a culture of each clone were analysed using SDS-PAGE. A discrete symbiont-specific band was detected in six clones. From the structural analyses of the insert in each clone, the candidate gene encoding the symbiont-specific protein was predicted and the amino acid sequence of the protein was deduced. The amino acid sequence in the N-terminal region of each protein was identical to that deduced from the genomic DNA sequence of the symbiont, but not of the host. The promoter sequences of the symbiont genes, very similar to those of the corresponding E. coli genes, were found in the insert DNA. These findings clearly indicate that genes derived from genomic DNA fragments of the P. stali symbiont can be expressed in E. coli.
authors.affiliation: Laboratory of Special Project, National Institute of Agrobiological Sciences, Ohwashi, Tsukuba, Ibaraki 305-8634, Japan. rkfujii@nias.affrc.go.jp
authors.forename: Rika
authors.initials: R
authors.lastname: Fujii-Muramatsu
authors.forename: Hideaki
authors.initials: H
authors.lastname: Kobayashi
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Noda
authors.forename: Keiichi
authors.initials: K
authors.lastname: Takeishi
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015698:Genomic Library; D020062:Heteroptera / Q000235:genetics / Q000382:microbiology*; D013559:Symbiosis
keywords: Escherichia coli; gene expression; genome; insect; symbiont