| 苏钟猪TLR4多态性及编码区C1027A功能分析 |
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| 引用本文： | 刘筱,方晓敏,邹晓龙,赵芳,任守文.苏钟猪TLR4多态性及编码区C1027A功能分析[J].中国农业科学,2012,45(6):1206-1214. |
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| 作者姓名： | 刘筱 方晓敏 邹晓龙 赵芳 任守文 |
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| 作者单位： | 1.江苏省农业科学院畜牧研究所，南京210014;
2.南京农业大学动物科学院，南京210095 |
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| 基金项目： | 国家生猪产业技术体系(nycytx-009);江苏省农业科技自主创新基金项目(CX(10)121);转基因生物新品种培育重大专项(2009ZX08009-153B);江苏省农科院学科带头人优秀后备人才项目(6510818);中波合作项目 |
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| 摘 要： | 【目的】研究苏钟猪TLR4的多态性及其编码区第1 027 bp处突变对TLR4蛋白功能的影响。【方法】采用PCR-SSCP的方法检测TLR4在苏钟猪中的多态性并利用real-time PCR方法检测不同基因型（CC型和AC型）的猪肺泡巨噬细胞经脂多糖（lipopolysaccharide，LPS）诱导后，TLR4及促炎症因子TNF-α、IL-1β表达量的变化情况，探讨该处突变对TLR4识别LPS的影响。【结果】①共检测到3个突变：G962A、C1027A、G960A，其中前两个为有义突变，且C1027A突变可引起编码氨基酸性质的改变；②LPS能快速诱导猪肺泡巨噬细胞中TLR4及促炎症因子TNF-α和IL-1β表达水平上调且TLR4编码区C1027A不同基因型（CC型和AC型）的猪肺泡巨噬细胞（pulmonary alveolar macrophages，PAMs）对LPS的敏感性及反应强度存在显著性差异。【结论】苏钟猪TLR4编码区的序列相对保守、多态含量较低，群体遗传变异程度较小。TLR4编码区C1027A突变能影响TLR4识别LPS的能力，等位基因C为苏钟猪抗革兰阴性菌感染的优势基因。
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| 关 键 词： | 苏钟猪 TLR4 PCR-SSCP LPS TNF-α IL-1&beta |
| 收稿时间： | 2011-03-04 |
Polymorphism of TLR4 and Function Analysis of C1027A in Suzhong Pig |
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| LIU Xiao
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FANG Xiao-min
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ZOU Xiao-long
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ZHAO Fang
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REN Shou-wen.Polymorphism of TLR4 and Function Analysis of C1027A in Suzhong Pig[J].Scientia Agricultura Sinica,2012,45(6):1206-1214. |
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| Authors: | LIU Xiao FANG Xiao-min ZOU Xiao-long ZHAO Fang REN Shou-wen |
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| Institution: | 1(1Institute of Animal Science,Jiangsu Academy of Agricultural Sciences,Nanjing 210014;2College of Animal Science and Technology Nanjing Agricultural University,Nanjing 210095) |
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| Abstract: | As a significant pattern-recognition receptor,toll-like receptor(TLR) is essential for the natural immunity.Its polymorphism has a profound influence on responses to a wide range of pathogens and is associated with resistance and susceptibility to diseases.【Objective】This study speculated on the polymorphisms of TLR4 and function analysis of the mutation(C1027A) in its coding sequence in Suzhong pig.【Method】 PCR-SSCP was used to search the mutations of TLR4 in 127 samples of Suzhong pigs.The expression levels of TLR4 and proinflammatory cytokines TNF-α and IL-1β were detected by real-time PCR in porcine alveolar macrophages(PAMs) that were induced by LPS.And the PAMs were grouped by their different genotypes(CC and AC) in 1027 bp of coding sequence(CDS) to discuss whether this mutation of TLR4 influenced its sensitivity to LPS.【Result】 Three SNPs were found,including G962A,C1027A and G960A,and the first two of them were nonsynonymous SNPs and C1027A changed the character of amino acid.LPS stimulation markedly increased TLR4 and proinflammatory cytokines TNF-α and IL-1β mRNA expression.And PAMs had different sensitivities and response to LPS between genotypes CC and AC of C1027A.【Conclusion】The coding sequence of TLR4 was relatively conservative in Suzhong pig and this population had low polymorphism and genetic variation.The mutation in C1027A can affect the binding ability to LPS.So,allele C in 1027 bp of TLR4 is the resistance allele to Gram-negative bacterial infections in Suzhong pig. |
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| Keywords: | Suzhong pig TLR4 PCR-SSCP LPS TNF-α IL-1β |
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