Skip to main content
SpringerLink
Log in

Spiralin-like protein SLP31 from Spiroplasma eriocheiris as a potential antigen for immunodiagnostics of tremor disease in Chinese mitten crab Eriocheir sinensis

  • Published:
Folia Microbiologica Aims and scope Submit manuscript

Abstract

Spiroplasma eriocheiris caused massive mortality of Chinese mitten crab Eriocheir sinensis but little is known about the molecular characteristics of this microorganism. We described here the identification of a spiralin-like protein (SLP31) from S. eriocheiris and expression in Escherichia coli. Analysis of the nucleotide sequence revealed that the clone has an open reading frame of 837 bp encoding a protein of 279 amino acids. Theoretical isoelectric point and molar mass for SLP31 are 7.72 and 31 kDa, respectively. The similarity of SLP31 deduced amino acid sequence shared with the spiralin from other species indicated that the gene may be a member of spiralin family. The TGA codon in Spiroplasma serves not as a stop signal but as a code for the amino acid tryptophan. After cloning the SLP31, the gene was site-mutated from TGA to TGG and transcribed in E. coli to full expression of SLP31. The purified recombinant protein was used to determine the immune reactivity by Western blotting which suggests that SLP31 could be a good antigen for immunodiagnostic of tremor disease in E. sinensis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BSA:

bovine serum albumin

DAB:

3,3′-diaminobenzidine tetrahydrochloride

HRP:

horseradish peroxidase

IPTG:

isopropyl thio-β-d-galactoside

Ni-NTA:

nickelnitrilotriacetate

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate-buffered saline

PCR:

polymerase chain reaction

PVDF:

polyvinylidene difluoride

SDS:

sodium dodecyl sulfate

S.e. :

Spiroplasma eriocheiris

SLP:

spiralin-like protein

TD:

tremor disease

References

  • Bi K., Huang H., Gu W., Wang J.H., Wang W.: Phylogenetic analysis of spiroplasmas from three freshwater crustaceans (Eriocheir sinensis, Procambarus clarkia and Penaeus vannamei) in China. J.Invertebr.Pathol.99, 57–65 (2008).

    Article  CAS  PubMed  Google Scholar 

  • Chambaud I., Wroblewski H., Blanchard A.: Interactions between mycoplasma lipoproteins and the host immune system. Trends Microbiol. 493–499 (1999).

  • Chevalier C., Saillard C., Bove J.M.: Spiralins of Spiroplasma citri and Spiroplasma melliferum: amino acid sequences and putative organization in the cell membrane. J.Bacteriol.172, 6090–6097 (1990).

    CAS  PubMed  Google Scholar 

  • Citti C., Marechal-drouard L., Saillard C., Weil J.H., Bove J.M.: Spiroplasma citri UGG and UGA tryptophan codons: sequence of the two tryptophanyl-tRNAs and organization of the corresponding genes. J.Bacteriol.174, 6471–6478 (1992).

    CAS  PubMed  Google Scholar 

  • Duret S., Berho N., Danet J.L., Garnier M., Renaudin J.: Spiralin is not essential for helicity, motility, or pathogenicity but is required for efficient transmission of Spiroplasma citri by its leafhopper vector Circulifer haematoceps. Appl.Environ.Microbiol.69, 6225–6234 (2003).

    Article  CAS  PubMed  Google Scholar 

  • Foissac X., Bove J.M., Saillard C.: Sequence analysis of Spiroplasma phoeniceum and Spiroplasma kunkelii spiralin genes and comparison with other spiralin genes. Curr.Microbiol.35, 240–243 (1997).

    Article  CAS  PubMed  Google Scholar 

  • Gasparich G.E., Whitcomb R.F., Dodge D., French F.E., Glass J., Williamson D.L.: The genus Spiroplasma and its non-helical descendants: phylogenetic classification, correlation with phenotype and roots of the mycoplasma mycoides clade. Internat. J.Syst.Evol.Microbiol.54, 893–918 (2004).

    Article  CAS  Google Scholar 

  • Killiny N., Castroviejo M., Saillard C.: Spiroplasma citri spiralin acts in vitro as a lectin binding to glycoproteins from its insect vector circulifer haematoceps. Phytopathology95, 541–548 (2005).

    Article  CAS  PubMed  Google Scholar 

  • Mouches C., Candresse T., Barroso G., Saillard C., Wroblewski H., Bove J.M.: Gene for spiralin, the major membrane protein of the helical mollicute Spiroplasma citri: cloning and expression in Escherichia coli. J.Bacteriol.164, 1094–1099 (1985).

    CAS  PubMed  Google Scholar 

  • Razin S., Yogev D., Naot Y.: Molecular biology and pathogenicity of mycoplasmas. Microbiol.Mol.Biol.Rev.62, 1094–1156 (1998).

    CAS  PubMed  Google Scholar 

  • Regassa L.B., Gasparich G.E.: Spiroplasmas: evolutionary relationships and biodiversity. Frontiers Biosci.11, 2983–3002 (2006).

    Article  CAS  Google Scholar 

  • Renaudin J., Marais A., Verdin E., Duret S., Foissac X., Laigret F., Bove J.M.: Integrative and free Spiroplasma citri oriC plasmids: expression of the Spiroplasma phoeniceum spiralin in Spiroplasma citri. J.Bacteriol.177, 2870–2877 (1995).

    CAS  PubMed  Google Scholar 

  • Stamburski C., Renaudin J., Bove J.M.: Mutagenesis of a tryptophan codon from TGG to TGA in the cat gene does not affect the expression of the cat gene in Spiroplasma citri. Gene110, 133–134 (1992).

    Article  CAS  PubMed  Google Scholar 

  • Wang J.H., Huang H., Feng Q., Liang T.M., Bi K.R., Gu W., Wang W., Shields J.D.: Enzyme-liked immunosorbent assay for the detection of pathogenic spiroplasma in commercially exploited crustaceans from China. Aquaculture292, 166–171 (2009).

    Article  CAS  Google Scholar 

  • Wang W., Zhu N.N., Gu Z.F., Du K., Xu Z.K.: Study on the transmission of tremor disease (TD) in the chinese mitten crab, Eriocheir sinensis (Crustacea:Decapoda). J.Invertebr.Pathol.81, 202–204 (2002).

    Article  PubMed  Google Scholar 

  • Wang W., Rong L.W., Gu W., Du K.H., Chen J.X.: Study on experimental infections of spiroplasma from the chinese mitten crab in crayfish, mice and embryonated chickens. Res.Microbiol.154, 677–680 (2003).

    Article  PubMed  Google Scholar 

  • Wang W., Chen J.X., Du K.H., Xu Z.K.: Morphology of spiroplasmas in the chinese mitten crab Eriocheir sinensis associated with tremor disease. Res.Microbiol.155, 630–635 (2004a).

    Article  CAS  PubMed  Google Scholar 

  • Wang W., Wen B.H., Gasparich G.E., Zhu N.N., Rong L.W., Chen J.X., Xu Z.K.: A spiroplasma associated with tremor disease in the chinese mitten crab (Eriocheir sinensis). Microbiology SGM150, 3035–3040 (2004b).

    Article  CAS  Google Scholar 

  • Wang W., Gu W., Ding Z.F., Ren Y.L., Chen J.X., Hou Y.Y.: A novel spiroplasma pathogen causing systemic infection in the crayfish Procambarus clarkii (Crustacea:Decapoda), in China. FEMS Microbiol.Lett.249, 131–137 (2005).

    Article  CAS  PubMed  Google Scholar 

  • Whitcomb R.F., Tully J.G., Wroblewski H.: Spiralin: major membrane protein specific for subgroup 1-1 spiroplasmas. Curr.Microbiol.9, 7–12 (1983).

    Article  Google Scholar 

  • Wroblewski H.: Electrophoretic analysis of the arrangement of spiralin and other major proteins in isolated Spiroplasma citri cell membranes. J.Bacteriol.145, 61–67 (1981).

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing, 210046, China

    Qingguo Meng, Wei Gu, Huyan Ji & Wen Wang

  2. Jiangsu Key Laboratory of Marine Biotechnology, Huaihai Institute of Technology, Lianyungang Jiangsu, 222005, China

    Keran Bi

  3. College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Qixia district, Nanjing, 210046, China

    Wen Wang

Authors
  1. Qingguo Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wei Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Keran Bi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Huyan Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wen Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wen Wang.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Meng, Q., Gu, W., Bi, K. et al. Spiralin-like protein SLP31 from Spiroplasma eriocheiris as a potential antigen for immunodiagnostics of tremor disease in Chinese mitten crab Eriocheir sinensis . Folia Microbiol 55, 245–250 (2010). https://doi.org/10.1007/s12223-010-0036-3

Download citation

  • Received:

  • Revised:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12223-010-0036-3

Keywords

Search

Navigation