猪TLR4基因外显子3的多态性研究

TLR4基因是识别革兰氏阴性菌内毒素-脂多糖（LPS）的主要受体，其多态性与动物对相关病原的抵抗力和易感性明显相关。本实验利用42个晋汾白猪和38个山西黑猪的基因组DNA，采用PCR-SSCP方法对TLR4基因外显子3的多态性进行分析。利用PopGene32软件对基因型检测结果进行群体遗传学分析，结果显示：晋汾白猪和山西黑猪的TLR4基因外显子3位点处均存在两种等位基因（T、A）和3种基因型（AA、TT和AT）。其中，在晋汾白猪中AA、AT、TT三种基因型频率分别为0.4524、0.2857和0.2619。山西黑猪中AA、AT、TT三种基因型频率分别为0.1579、0.2105和0.6316。晋汾白猪TLR4基因的纯合度为0.7143，香农指数为0.6749；山西黑猪TLR4基因的纯合度为0.7895，香农指数为0.5763。晋汾白猪和山西黑猪在TLR4基因外显子3上的基因型分布均不符合哈代-温伯格平衡（p<0.05）。利用SAS8.1统计软件进行卡方检验表明，晋汾白猪和山西黑猪的基因型分布在TLR4基因的多态位点差异不显著（p>0.05）。通过对晋汾白猪和山西黑猪TLR4基因外显子3的多态性分析，能够为晋汾白猪和山西黑猪的抗病育种提供遗传学依据。

关键词：TLR4基因；PCR-SSCP；多态性；晋汾白猪；山西黑猪

The polymorphism detection of TLR4 gene exon 3 in pig
 

Abstract: TLR4 gene is the main receptor to identify the endotoxin - lipopolysaccharide ( LPS ) of gram - negative bacteria, and its polymorphism is related to the resistance and susceptibility of animals to related pathogens. Using the genomic DNA of 42 Jinfen white pigs and 38 Shanxi black pigs, the polymorphism of the exon 3 of TLR4 gene was analyzed by PCR - SSCP method. Using the software of pop gene 32, the genetic analysis of genotype was analyzed. The results showed that there were two alleles ( A, T ) and three genotypes ( AA, AT and TT ) at the 3 loci of TLR4 gene in Jinfen white pig and Shanxi black pig. Among them, the three genotypes of AA, AT and TT were 0.4524, 0.2857 and 0.2619 respectively in Jinfen white pig. The frequencies of AA, AT and TT in Shanxi black pigs were 0.1579, 0.2105 and 0.6316 respectively. The homozygosity of TLR4 gene in Jinfen white pig was 0.7143, and the Shannon index was 0.6749. The homozygosity of TLR4 gene in Shanxi black pig was 0.7895, and the Shannon index was 0.5763. The genotype distribution of Jinfen white pig and Shanxi black pig in exon 3 of TLR4 gene was not consistent with hardy - Weinberg equilibrium ( p < 0.05 ). Chi - square test using SAS 8.1 statistical software showed that the genotype distribution of Jinfen white pig and Shanxi black pig was not significantly different in the polymorphism site of TLR4( p > 0.05 ). Through the analysis of the polymorphism of the exon 3 of TLR4 gene in Jinfen white pig and Shanxi black pig, which provides relevant basis for the disease resistance breeding of Jinfen white pig and Shanxi black pig.

Key words: TLR4 gene; PCR-SSCP; Polymorphism; Jinfen white pig; Shanxi black pig

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