猪Toll样受体4基因SNPs功能分析

doi:10.3724/SP.J.1005.2012.01050

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HEREDITAS ›› 2012, Vol. 34 ›› Issue (8): 1050-1056.doi: 10.3724/SP.J.1005.2012.01050

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Functional analysis of SNPs in porcine TLR4 gene

YANG Xiu-Qin¹, CHEN Yue-Chan¹, WANG Li-ang², LI Hai-Tao¹, LIU Di², GUAN Qing-Zhi¹, FU Bo²

1. College of Animal Science & Technology, Northeast Agricultural University, Harbin 150030, China 2. Heilongjiang Academy of Agricultural Science, Harbin 150086, China

Received:2012-01-03 Revised:2012-03-15 Online:2012-08-20 Published:2012-08-25

Abstract

Abstract: Toll-like receptor 4 (TLR4) plays an important role in immune response and the polymorphism in it might affect protein signaling and host resistance/susceptibility to disease. This study was designed to characterize the functional relevance of 3 nonsynonymous single nucleotide polymorphisms (SNPs), c.611 T>A (p.Leu204His), c.1027 C>A (p.Gln343Lys), and c.1605 G>T (p.Leu535Phe), which were selected based on our previous studies. RT-PCR method was used to clone the complete coding sequence of porcine TLR4 gene and the PCR-based method was used to introduce the point mutation. The effects of 3 SNPs on the ligand recognition and signaling of porcine TLR4 were investigated in transiently transfected PK-15 cells using dual-luciferase reporter system and Western blotting method. At the same time, the distribution of c.1605 G>T among pig populations composed of Min pig, Yorkshire, Landrace, and Wild boar from north-eastern China was studied by created restriction site PCR-RFLP method. The complete coding sequence of TLR4 gene in Min pig and 3 variants with single point mutations were obtained. Eukaryotic expression vectors containing differ-ent alleles of porcine TLR4 were constructed. SNP c.1605 G>T significantly decreased the TLR4 signaling (P<0.01) and the polymorphism only existed in Min pig and Wild boar from northeastern China with high frequencies. SNP c.1605 G>T in porcine TLR4 might affect the receptor function and host resistance/susceptibility to diseases.

Key words: porcine, TLR4, SNP, functional analysis

YANG Xiu-Qin, CHEN Yue-Chan, WANG Li-ang, LI Hai-Tao, LIU Di, GUAN Qing-Zhi, FU Bo. Functional analysis of SNPs in porcine TLR4 gene[J]. HEREDITAS, 2012, 34(8): 1050-1056.

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Functional analysis of SNPs in porcine TLR4 gene

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