Format

Send to:

Choose Destination
See comment in PubMed Commons below
Fish Shellfish Immunol. 2013 Sep;35(3):653-60. doi: 10.1016/j.fsi.2013.05.024. Epub 2013 May 31.

Molecular cloning and expression analysis of polymeric immunoglobulin receptor in flounder (Paralichthys olivaceus).

Author information

  • 1Laboratory of Pathology and Immunology of Aquatic Animals, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China.

Abstract

The polymeric immunoglobulin receptor (pIgR) is one of the most important mucosal effectors mediating the transcytosis of polymeric immunoglobulins (pIgs) to protect the organisms. In this paper, a full-length cDNA of pIgR was firstly cloned from flounder (Paralichthys olivaceus) using rapid amplification of cDNA ends approaches, and it was of 1384 bp, containing an open reading frame (ORF) of 1005 bp encoding a polypeptide of 335 amino acids with the predicted molecular mass of 37.6 kDa. The flounder pIgR exhibited a unique structure containing only two immunoglobulin-like domains (ILD) corresponding to mammalian pIgR ILD1 and ILD5. The mRNA transcripts of pIgR were detected in all the tested tissues of flounder by semi-quantitative RT-PCR, and the pIgR was expressed at the highest level in liver and higher levels in intestine, gill, skin, spleen and head kidney than in stomach and muscle. The ORF was successfully expressed in Escherichia coli BL21 (DE3) and the recombinant protein displayed binding capability to the purified mucus IgM and serum IgM of flounder by ELISA. The polyclonal antibody against flounder recombinant pIgR was developed by immunization of Balb/C mice, which specifically reacted to the recombinant pIgR in Western blot. Moreover, a secretory component-like molecule was detected in the skin mucus but not in the serum of flounder, which molecular mass (about 37 kDa) was near the theoretical mass obtained from the sequence of flounder pIgR. All these results indicated that flounder pIgR probably involved in the pIgs transport and provided insights into the roles of fish pIgR in the mucosal immunity.

Copyright © 2013 Elsevier Ltd. All rights reserved.

KEYWORDS:

Flounder (Paralichthys olivaceus); Immunoglobulin; Mucosal immunity; Polymeric immunoglobulin receptor (pIgR); RT-PCR

PMID:
23732846
[PubMed - indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Write to the Help Desk