title: Transformation of crustacean pathogenic bacterium Spiroplasma eriocheiris and expression of yellow fluorescent protein.
abstract: Spiroplasma eriocheiris, the cause of crab trembling disease, is a wall-less bacterium, related to Mycoplasmas, measuring 2.0-10.0 μm long. It features a helical cell shape and a unique swimming mechanism that does not use flagella; instead, it moves by switching the cell helicity at a kink traveling from the front to the tail. S. eriocheiris seems to use a novel chemotactic system that is based on the frequency of reversal swimming behaviors rather than the conventional two-component system, which is generally essential for bacterial chemotaxis. To identify the genes involved in these novel mechanisms, we developed a transformation system by using oriC plasmid harboring the tetracycline resistant gene, tetM, which is under the control of a strong promoter for an abundant protein, elongation factor-Tu. The transformation efficiency achieved was 1.6 × 10-5colony forming unit (CFU) for 1 μg DNA, enabling the expression of the enhanced yellow fluorescent protein (EYFP).
authors.affiliation: Department of Biology, Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan. Electronic address: natsuholy@gmail.com.
authors.forename: Natsuho
authors.initials: N
authors.lastname: Terahara
authors.affiliation: Department of Biology, Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan; The OCU Advanced Research Institute for Natural Science and Technology (OCARINA), Osaka City University, Sumiyoshi-ku, Osaka 558-8585, Japan. Electronic address: iciltlm@gmail.com.
authors.forename: Isil
authors.initials: I
authors.lastname: Tulum
authors.affiliation: Department of Biology, Graduate School of Science, Osaka City University, 3-3-138 Sugimoto, Sumiyoshi-ku, Osaka 558-8585, Japan; The OCU Advanced Research Institute for Natural Science and Technology (OCARINA), Osaka City University, Sumiyoshi-ku, Osaka 558-8585, Japan. Electronic address: miyata@sci.osaka-cu.ac.jp.
authors.forename: Makoto
authors.initials: M
authors.lastname: Miyata
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008164:Luminescent Proteins / Q000235:genetics*; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D013149:Spiroplasma / Q000235:genetics*; D014169:Transformation, Bacterial
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria
keywords: EYFP; Mollicutes; Mycoplasma; Spiroplasma; Swimming; oriC plasmid