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断奶仔猪TLR4基因多态性及组织表达分析

Analysis of TLR4 Gene Polymorphism and Tissue Expression in Weaned Piglets

农业科学进展 / 2019,1(1)：1-14 / 2019-09-11 6251 9233

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• 作者： 刘晓慧¹²      冯巩俐¹      张雅君¹      孙奴奴¹*     
• 单位：

  1.运城学院生命科学系，运城；
  2.上海海洋大学，上海

• 关键词： TLR4 基因；晋汾白猪；新山西黑猪；RT-PCR；PCR-SSCP；多态性；组织表达
• TLR4 gene; Jinfen white pig; new Shanxi black pig; RT-PCR; PCR-SSCP; Polymorphism; Tissue expression
• 摘要： TLR4基因是脂多糖（LPS）的主要受体，连接机体的代谢和免疫调控。本研究采用 PCR-SSCP 技术，对猪的TLR4基因外显子1和3的单核苷酸多态性进行分析，结果表明：晋汾白猪和新山西黑猪的TLR4基因外显子1处均存在4种基因型（AA、BB、AB、CC），在外显子3处均存在3种基因型（AA、BB、AB），卡方检验表明，两个猪种的基因型分布在TLR4基因的多态位点差异不显著（p>0.05）。采用RT-PCR技术，检测TLR4基因在晋汾白猪和新山西黑猪不同组织中的表达水平，结果显示：TLR4基因在晋汾白猪和新山西黑猪各个组织中都表达。应用SPSS 19.0分析TLR4基因在两个猪种的四种免疫组织（脾、巨噬细胞、白细胞、淋巴结）间的表达差异情况。结果表明：TLR4基因的表达在晋汾白猪的脾与白细胞、巨噬细胞、淋巴结间差异非常显著（p<0.01）；在新山西黑猪的淋巴结与白细胞、巨噬细胞、脾间差异非常显著（p<0.01）；在两个猪种的脾脏组织间差异非常显著（p<0.01）。研究成功建立了TLR4基因变异信息及其组织表达情况，将为进一步分析猪的TLR4基因变异与猪的抗病育种提供基础资料。
• TLR4 gene is the main receptor for lipopolysaccharide (LPS), which connects the body's metabolism and immune regulation. In this study, the single nucleotide polymorphisms of exons 1 and 3 of TLR4 gene in pigs was detected by PCR-SSCP. The results showed that there were 4 genotypes (AA, BB, AB, CC) in exon 1 of TLR4 gene in Jinfen white pig and new Shanxi black pig, and 3 genotypes (AA, BB, AB) in exon 3. The Chi-square test showed that the genotype distribution was not significantly difference between Jinfen white pig and new Shanxi black pig at the polymorphic sites of TLR4 gene (p>0.05). In addition, the expression level of TLR4 gene in different tissues was detected by RT-PCR. The results showed that TLR4 gene was expressed in every tissue of Jinfen white pig and new Shanxi black pig respectively. SPSS 19.0 software was used to further analyze the expression difference of TLR4 gene between four kinds of immune tissues (spleen, macrophage, leukocyte and lymph node) of the two pig species. The results showed that the expression of TLR4 gene was significantly different between the spleen of Jinfen white pig and leukocytes, macrophages, lymph nodes (p<0.01). There were extremely significant differences in the expression between the lymph nodes and white blood cells, macrophages, and the spleen of the new Shanxi black pig (p<0.01). The expression difference between spleens of Jinfen white pig and new Shanxi black pig was extremely significant (p<0.01). The experiment successfully established TLR4 gene mutation information and tissue expression profile, which would provide basic data for further analysis of TLR4 gene variation and breeding of disease resistance in pigs.
• DOI： https://doi.org/10.35534/aas.0101001
• 引用： 刘晓慧，冯巩俐，张雅君，等．断奶仔猪TLR4基因多态性及组织表达分析［J］.农业科学进展，2019，1（1）：1-14.
  https://doi.org/10.35534/aas.0101001