Skip to main content
Log in

Cloning and expression pattern of a hemolin homologue from the diamondback moth, Plutella xylostella

  • Research Article
  • Published:
Genes & Genomics Aims and scope Submit manuscript

Abstract

Hemolin has been known to play a key role in insect innate immunity. In an attempt to examine expression pattern of the Hemolin gene in the diamondback moth, Plutellea xylostella, the full-length cDNA of Hemolin was cloned using 5′-RACE PCR technique. The cDNA contained a 5′ untranslated region of 48 nucleotides and a 3′ untranslated region of 198 nucleotides, including a stop codon (TAA) and a poly (A) tail. It consists of 1,401 bp with an open reading frame of 1,245 bp, encoding 414 amino acids. The deduced amino acid sequence of PxHemolin has relatively low identities (35−42%) to various insect Hemolins. However, it has high three-dimensional structural similarity to Hemolin. Interestingly, analysis of spatial expression pattern of PxHemolin shows that it was highly expressed in the Malpighian tubule and the silk gland although it was also detected in fat body and gut. Furthermore, PxHemolin mRNA was highly induced 3 hr after immune-challenging with lipopolysaccharide and was gradually up-regulated after laminarin treatment. These data suggest that PxHemolin may play a role in innate immune responses although it remains to further elucidate the precise biological functions in P. xylostella.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bao Y, Yamano Y and Morishima I (2007) Induction of hemolin gene expression by bacterial cell wall components in eri-silkworm, Samia cynthia ricini. Comp. Biochem. Physiol. B. Biochem. Mol. Biol. 146: 147–151.

    Article  PubMed  Google Scholar 

  • Bates PA, Kelley LA, MacCallum RM and Sternberg MJ (2001) Enhancement of protein modeling by human intervention in applying the automatic programs 3D-JIGSAW and 3D-PSSM. Proteins Suppl. 5: 39–46.

    Article  Google Scholar 

  • Bates PA and Sternberg MJ (1999) Model building by comparison at CASP3: using expert knowledge and computer automation. Proteins Suppl. 3: 47–54.

    Article  Google Scholar 

  • Bendtsen JD, Nielsen H, von Heijne G and Brunak S (2004) Improred prediction of signal peptides: Signal IP 3.0. J. Mol. Biol. 340: 783–795.

    Article  PubMed  Google Scholar 

  • Bettencourt R, Assefaw-Redda Y and Faye I (2000) The insect immune protein hemolin is expressed during oogenesis and embryogenesis. Mech. Dev. 95: 301–304.

    Article  CAS  PubMed  Google Scholar 

  • Contreras-Moreira B and Bates PA (2002) Domain fishing: a first step in protein comparative modelling. Bioinformatics 18: 1141–1142.

    Article  CAS  PubMed  Google Scholar 

  • Daffre S and Faye I (1997) Lipopolysaccharide interaction with hemolin, an insect member of the Ig-superfamily. FEBS Lett. 408: 127–130.

    Article  CAS  PubMed  Google Scholar 

  • Eleftherianos I, Gokcen F, Felfoldi G, Millichap PJ, Trenczek TE, ffrench-Constant RH and Reynolds SE (2007) The immunoglobulin family protein Hemolin mediates cellular immune responses to bacteria in the insect Manduca sexta. Cell Microbiol. 9: 1137–1147.

    Article  CAS  PubMed  Google Scholar 

  • Eleftherianos I, Millichap PJ, ffrench-Constant RH and Reynolds SE (2006) RNAi suppression of recognition protein mediated immune responses in the tobacco hornworm Manduca sexta causes increased susceptibility to the insect pathogen Photorhabdus. Dev. Comp. Immunol. 30: 1099–1107.

    Article  CAS  PubMed  Google Scholar 

  • Hunter S, Apweiler R, Attwood TK, Bairoch A, Bateman A, Binns D, Bork P, Das U, Daugherty L, Duquenne L, Finn RD, et al. (2009) InterPro: the integrative protein signature database. Nucleic Acids Res. 37 (Database issue): D211–215.

    Article  CAS  PubMed  Google Scholar 

  • Lanz-Mendoza H, Bettencourt R, Fabbri M and Faye I (1996) Regulation of the insect immune response: the effect of hemolin on cellular immune mechanisms. Cell Immunol. 169: 47–54.

    Article  CAS  PubMed  Google Scholar 

  • Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, et al. (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23: 2947–2948.

    Article  CAS  PubMed  Google Scholar 

  • Li W, Terenius O, Hirai M, Nilsson AS and Faye I (2005) Cloning, expression and phylogenetic analysis of Hemolin, from the Chinese oak silkmoth, Antheraea pernyi. Dev. Comp. Immunol. 29: 853–864.

    Article  CAS  PubMed  Google Scholar 

  • Lindstrom-Dinnetz I, Sun SC and Faye I (1995) Structure and expression of Hemolin, an insect member of the immunoglobulin gene superfamily. Eur. J. Biochem. 230: 920–925.

    Article  CAS  PubMed  Google Scholar 

  • Noh M, Jo Y, Oh S, Kim D, Park H, Kim I, Barillas-Mury C, Kim H, Lee W, Lee I, et al. (2006) Cloning and subcellular localization of a serpin containing nuclear export signal from the Korean malaria vector, Anopheles sinensis. Kor. J. Genet. 28: 433–441.

    CAS  Google Scholar 

  • Noh M, Kim H, Jo Y, Park C, Park H, Kim S, Park J, Kim I, Seo S, Lee Y, et al. (2007) Molecular cloning and expression pattern of a serpin-2 from the beet armyworn, Spodoptera exigua. Kor. J. Genet. 29: 255–261.

    CAS  Google Scholar 

  • Saitou N and Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4: 406–425.

    CAS  PubMed  Google Scholar 

  • Shaik HA and Sehnal F (2009) Hemolin expression in the silk glands of Galleria mellonella in response to bacterial challenge and prior to cell disintegration. J. Insect Physiol. 55: 781–787.

    Article  CAS  PubMed  Google Scholar 

  • Shin SW, Park SS, Park DS, Kim MG, Kim SC, Brey PT and Park HY (1998) Isolation and characterization of immune-related genes from the fall webworm, Hyphantria cunea, using PCR-based differential display and subtractive cloning. Insect Biochem. Mol. Biol. 28: 827–837.

    Article  CAS  PubMed  Google Scholar 

  • Su XD, Gastinel LN, Vaughn DE, Faye I, Poon P and Bjorkman PJ (1998) Crystal structure of hemolin: a horseshoe shape with implications for homophilic adhesion. Science 281: 991–995.

    Article  CAS  PubMed  Google Scholar 

  • Sun SC, Lindstrom I, Boman HG, Faye I and Schmidt O (1990) Hemolin: an insect-immune protein belonging to the immunoglobulin superfamily. Science 250: 1729–1732.

    Article  CAS  PubMed  Google Scholar 

  • Tamura K, Dudley J, Nei M and Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol. Biol. Evol. 24: 1596–1599.

    Article  CAS  PubMed  Google Scholar 

  • Terenius O (2008) Hemolin-A lepidopteran anti-viral defense factor? Dev. Comp. Immunol. 32: 311–316.

    Article  CAS  PubMed  Google Scholar 

  • Terenius O, Popham HJ and Shelby KS (2009) Bacterial, but not baculoviral infections stimulate Hemolin expression in noctuid moths. Dev. Comp. Immunol. 33: 1176–1185.

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Willott E and Kanost MR (1995) Organization and expression of the hemolin gene, a member of the immunoglobulin superfamily in an insect, Manduca sexta. Insect Mol. Biol. 4: 113–123.

    Article  CAS  PubMed  Google Scholar 

  • Yu XQ and Kanost MR (1999) Developmental expression of Manduca sexta hemolin. Arch Insect Biochem. Physiol. 42: 198–212.

    Article  CAS  PubMed  Google Scholar 

  • Yu XQ and Kanost MR (2002) Binding of hemolin to bacterial lipopolysaccharide and lipoteichoic acid. An immunoglobulin superfamily member from insects as a pattern-recognition receptor. Eur. J. Biochem. 269: 1827–1834.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Yeon Soo Han.

Additional information

R. J. Kim and Y. H. Jo are equally contributed to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, R.J., Jo, Y.H., Oh, S.H. et al. Cloning and expression pattern of a hemolin homologue from the diamondback moth, Plutella xylostella . Genes Genom 32, 71–77 (2010). https://doi.org/10.1007/s13258-010-0834-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13258-010-0834-7

Keywords

Navigation