Molecular cloning and functional analysis of duck Toll-like receptor 5

Res Vet Sci. 2014 Aug;97(1):43-5. doi: 10.1016/j.rvsc.2014.05.010. Epub 2014 Jun 11.

Abstract

Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In this study, we cloned the single-exon TLR5 gene of the Maya breed of Common Shelduck (Tadorna tadorna). The TLR5 open reading frame is 2580 bp in length and encodes an 859-amino acid protein. The putative amino acid sequence of duck TLR5 consisted of a signal peptide sequence, 11 leucine-rich repeat domains, a leucine-rich repeat C-terminal domain, a transmembrane domain, and an intracellular Toll-interleukin-1 receptor domain. The duck TLR5 gene was highly expressed in the lung, bone marrow, spleen, and liver; moderately expressed in kidney, small intestine, large intestine, and brain. A plasmid expressing duck TLR5 was constructed and transfected into HEK293T cells, and expression was confirmed by indirect immunofluorescence assay. HEK293T cells transfected with duck TLR5- and NF-κB-luciferase-containing plasmids significantly responded to flagellin from Salmonella typhimurium, indicating that it is a functional TLR5 homolog.

Keywords: Duck; Expression; Flagellin; Function; Molecular cloning; Toll-like receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cloning, Molecular*
  • Ducks / genetics*
  • Ducks / physiology*
  • Flagellin / metabolism
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • Open Reading Frames / genetics
  • Plasmids / genetics
  • Salmonella typhimurium / metabolism
  • Toll-Like Receptor 5 / analysis
  • Toll-Like Receptor 5 / genetics*
  • Toll-Like Receptor 5 / physiology*
  • Transfection

Substances

  • Toll-Like Receptor 5
  • Flagellin