猪IRS4基因多态性、表达差异及其与脂肪沉积性状的关联分析

doi:10.11843/j.issn.0366-6964.2015.07.002

摘要/Abstract

摘要：

本研究旨在检测猪IRS4基因多态性及其在17种组织中的表达水平，寻找基因内与脂肪沉积性状QTL效应相关的分子标记。本研究采用比较测序方法搜寻到IRS4基因的3个SNPs，分别是在启动子上的g.96C>G，外显子上的同义突变g.1829T>C和错义突变g.1794T>C（p.Phe432Leu）。RT-PCR检测后发现，IRS4基因在垂体、下丘脑中高度表达，在其他组织中弱表达或不表达。在白色杜洛克×二花脸F₂资源家系、苏太猪群体、二花脸群体、杜长大三元杂商品猪及中国地方野猪中，用PCR-RFLP方法对IRS4的3个SNPs位点进行判型，然后分析它们的基因频率及其与4点（肩、胸、腰、臀部）背膘厚、腹脂重、肌内脂肪含量和眼肌面积的相关性。结果发现，g.96C>G位点在F₂家系（含有SSCX QTL）中分离程度很低，被首先排除作为潜在因果突变（QTN）的可能性。g.1794T>C和g.1829T>C位点在所有检测的西方猪种（白色杜洛克和杜长大）是均为TT基因型，而它们的CC基因型频率在二花脸猪中分别为65%和100%。在不同群体中标记关联分析结果显示，g.1794T>C与F₂个体所有脂肪沉积表型均极显著相关（P<0.01），但它在苏太猪中仅与IMF显著相关（P<0.05），且与二花脸猪表型关联不显著（P>0.05）；g.1829T>C则不仅与F2个体所有脂肪沉积表型均极显著相关（P<0.01），而且与苏太猪的4点膘厚和眼肌面积呈显著或极显著相关。此外，生物信息学分析发现，g.1794T>C错义突变可能对IRS4蛋白的功能影响不大。结果提示，该错义突变也不太可能是影响脂肪沉积的QTN，而g.1829T>C与F₂和苏太猪的脂肪沉积性状都显著关联，故其可作为这些性状选育的分子标记，并值得深入研究。

Abstract:

This study aimed to detect polymorphisms of porcine IRS4 gene and its expression pattern in 17 tissues，and to identify gene markers associated with the QTL for fat deposition traits.Comparative sequence analysis of the IRS4 revealed 3 SNPs，including g.96C>G in the promoter region，a synonymous substitution g.1829T>C and a missense substitution g.1794T>C (p.Phe432Leu).Through RT-PCR assay，we found that IRS4 gene was abundantly expressed in hypothalamus and pituitary，but weakly expressed in other tissues.Furthermore，we genotyped the 3 SNPs in White Duroc×Erhualian F₂ intercross，Sutai pigs，Erhualian pigs and DLY［Duroc×(Landrace×Yorkshire)］three-way crossbred pigs and Chinese local wild boar by PCR-RFLP，then calculated allele frequencies and estimated the associations between those SNPs and backfat thicknesses (BFTs) at shoulder，chest，waist and hip，abdominal fat weight (AFW)，intramuscular fat content (IMF) and loin eye area (LEA) in these populations.The SNP g.96C>G was almost fixed in the F₂ intercross，so it was first excluded as a causative mutation (QTN) underlying this QTL.For both SNPs g.1794T>C and g.1829T>C，all western breeds (White Duroc and DLY) only had TT genotype，while，in the Erhualian population，the frequencies of CC genotypes at g.1794T>C and g.1829T>C were 65% and 100%，respectively.The results of association analysis showed that g.1794T>C was highly associated with all investigated traits in the F₂ intercross (P<0.01)，but associated only with IMF in Sutai pigs（P<0.05）and had no significant effect on Erhualian’s phenotypes；g.1829T>C was significantly associated with several fatness traits (e.g.BFT and LEA) in the F₂ intercross and Sutai pigs.Moreover，bioinformatics analysis demonstrated that the potential influence of the missense variant g.1794T>C on IRS4 protein function was limited.The results suggest that the SNP g.1794T>C is unlikely to be QTN，and the SNP g.1829T>C associated with fat deposition traits across populations could be used as marker for selection of these traits.

中图分类号:

S828.2

马焕班，刘先先，杨杰，黄卫兵，段艳宇，郭源梅，麻骏武. 猪IRS4基因多态性、表达差异及其与脂肪沉积性状的关联分析[J]. 畜牧兽医学报, 2015, 46(7): 1084-1094.

MA Huan-ban，LIU Xian-xian，YANG Jie，HUANG Wei-bing，DUAN Yan-yu，GUO Yuan-mei，MA Jun-wu. Porcine IRS4 Gene：Polymorphism，Differential Expression and Its Associations with Fat Deposition Traits[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(7): 1084-1094.

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