Cargando…

Molecular Cloning and Functional Analysis of the Duck TLR4 Gene

Toll-like receptor 4 (TLR4) recognizes pathogen-associated molecular patterns in some animals and has been shown to be closely associated with several diseases such as tumors, atherosclerosis, and asthma. However, its function in ducks is not clear. Alternative splicing of the TLR4 gene has been ide...

Descripción completa

Detalles Bibliográficos
Autores principales: Zhao, Wenming, Huang, Zhengyang, Chen, Yang, Zhang, Yang, Rong, Guanghui, Mu, Chunyu, Xu, Qi, Chen, Guohong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3794798/
https://www.ncbi.nlm.nih.gov/pubmed/24025421
http://dx.doi.org/10.3390/ijms140918615
_version_ 1782287260074377216
author Zhao, Wenming
Huang, Zhengyang
Chen, Yang
Zhang, Yang
Rong, Guanghui
Mu, Chunyu
Xu, Qi
Chen, Guohong
author_facet Zhao, Wenming
Huang, Zhengyang
Chen, Yang
Zhang, Yang
Rong, Guanghui
Mu, Chunyu
Xu, Qi
Chen, Guohong
author_sort Zhao, Wenming
collection PubMed
description Toll-like receptor 4 (TLR4) recognizes pathogen-associated molecular patterns in some animals and has been shown to be closely associated with several diseases such as tumors, atherosclerosis, and asthma. However, its function in ducks is not clear. Alternative splicing of the TLR4 gene has been identified in pigs, sheep, mice, and other species, but has not yet been reported in the duck. In this study, alternative splicing of the duck TLR4 gene was investigated using reverse transcription-polymerase chain reaction (RT-PCR). Duck TLR4 gene (duTLR4, accession number: KF278109) was found to consist of 3367 nucleotides of coding sequence. An alternative splice form, TLR4-b, was identified and shown by alignment to retain the intron between exons 1 and 2. Real-time quantitative polymerase chain reaction (qPCR) analyses suggested that duTLR4-a (wild-type) mRNA is widely expressed in various healthy tissues, whereas TLR4-b is expressed at only low levels. Following stimulation of normal duck embryo fibroblasts with lipopolysaccharide, the expression of both isoforms initially increased and then decreased. Expression of the wild-type isoform subsequently increased again, while that of the variant remained low. The expression levels of wild-type TLR4 were further analyzed by transient transfection of a pcDNA3.1(+)-TLR4-a overexpression vector into duck embryo fibroblasts. qRT-PCR analyses showed that after stimulation with LPS and poly(I:C) the expression levels of IL-1β, IL6, and MHC II increased with a response-efficacy relationship. Our experimental results indicate that TLR4 plays an important role in resistance to both bacterial and viral infections in the duck.
format Online
Article
Text
id pubmed-3794798
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-37947982013-10-21 Molecular Cloning and Functional Analysis of the Duck TLR4 Gene Zhao, Wenming Huang, Zhengyang Chen, Yang Zhang, Yang Rong, Guanghui Mu, Chunyu Xu, Qi Chen, Guohong Int J Mol Sci Article Toll-like receptor 4 (TLR4) recognizes pathogen-associated molecular patterns in some animals and has been shown to be closely associated with several diseases such as tumors, atherosclerosis, and asthma. However, its function in ducks is not clear. Alternative splicing of the TLR4 gene has been identified in pigs, sheep, mice, and other species, but has not yet been reported in the duck. In this study, alternative splicing of the duck TLR4 gene was investigated using reverse transcription-polymerase chain reaction (RT-PCR). Duck TLR4 gene (duTLR4, accession number: KF278109) was found to consist of 3367 nucleotides of coding sequence. An alternative splice form, TLR4-b, was identified and shown by alignment to retain the intron between exons 1 and 2. Real-time quantitative polymerase chain reaction (qPCR) analyses suggested that duTLR4-a (wild-type) mRNA is widely expressed in various healthy tissues, whereas TLR4-b is expressed at only low levels. Following stimulation of normal duck embryo fibroblasts with lipopolysaccharide, the expression of both isoforms initially increased and then decreased. Expression of the wild-type isoform subsequently increased again, while that of the variant remained low. The expression levels of wild-type TLR4 were further analyzed by transient transfection of a pcDNA3.1(+)-TLR4-a overexpression vector into duck embryo fibroblasts. qRT-PCR analyses showed that after stimulation with LPS and poly(I:C) the expression levels of IL-1β, IL6, and MHC II increased with a response-efficacy relationship. Our experimental results indicate that TLR4 plays an important role in resistance to both bacterial and viral infections in the duck. MDPI 2013-09-10 /pmc/articles/PMC3794798/ /pubmed/24025421 http://dx.doi.org/10.3390/ijms140918615 Text en © 2013 by the authors; licensee MDPI, Basel, Switzerland http://creativecommons.org/licenses/by/3.0 This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Zhao, Wenming
Huang, Zhengyang
Chen, Yang
Zhang, Yang
Rong, Guanghui
Mu, Chunyu
Xu, Qi
Chen, Guohong
Molecular Cloning and Functional Analysis of the Duck TLR4 Gene
title Molecular Cloning and Functional Analysis of the Duck TLR4 Gene
title_full Molecular Cloning and Functional Analysis of the Duck TLR4 Gene
title_fullStr Molecular Cloning and Functional Analysis of the Duck TLR4 Gene
title_full_unstemmed Molecular Cloning and Functional Analysis of the Duck TLR4 Gene
title_short Molecular Cloning and Functional Analysis of the Duck TLR4 Gene
title_sort molecular cloning and functional analysis of the duck tlr4 gene
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3794798/
https://www.ncbi.nlm.nih.gov/pubmed/24025421
http://dx.doi.org/10.3390/ijms140918615
work_keys_str_mv AT zhaowenming molecularcloningandfunctionalanalysisoftheducktlr4gene
AT huangzhengyang molecularcloningandfunctionalanalysisoftheducktlr4gene
AT chenyang molecularcloningandfunctionalanalysisoftheducktlr4gene
AT zhangyang molecularcloningandfunctionalanalysisoftheducktlr4gene
AT rongguanghui molecularcloningandfunctionalanalysisoftheducktlr4gene
AT muchunyu molecularcloningandfunctionalanalysisoftheducktlr4gene
AT xuqi molecularcloningandfunctionalanalysisoftheducktlr4gene
AT chenguohong molecularcloningandfunctionalanalysisoftheducktlr4gene