Transformation of crustacean pathogenic bacterium Spiroplasma eriocheiris and expression of yellow fluorescent protein

Terahara N., Tulum I., Miyata M.

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, vol.487, no.3, pp.488-493, 2017 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 487 Issue: 3
  • Publication Date: 2017
  • Doi Number: 10.1016/j.bbrc.2017.03.144
  • Journal Name: BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.488-493
  • Keywords: Spiroplasma, oriC plasmid, Mollicutes, Mycoplasma, EYFP, Swimming, MYCOPLASMA-PNEUMONIAE, MELLIFERUM BC3, ORIC PLASMIDS, REPLICATION, IDENTIFICATION, LOCALIZATION, CHROMOSOME, MOLLICUTES, ORIGIN, CITRI
  • Istanbul University Affiliated: No

Abstract

Spiroplasma eriocheiris, the cause of crab trembling disease, is a wall-less bacterium, related to Mycoplasmas, measuring 2.0-10.0 mu m 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 x 10(-5) colony forming unit (CFU) for 1 mu g DNA, enabling the expression of the enhanced yellow fluorescent protein (EYFP). (C) 2017 The Authors. Published by Elsevier Inc.