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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2013
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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 |
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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 |
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