不同品种猪HSL基因 5′-UTR和外显子Ⅰ片段的克隆测序及多态性分析

Sequence and Polymorphism Analysis of Porcine Hormone-sensitive Lipase Gene 5′-UTR and Exon Ⅰ

激素敏感脂肪酶(Hormone sensitivelipase,HSL)是负责分解脂肪组织中甘油三酯释放游离脂肪酸的关键和限速酶,也是影响动物脂肪沉积的关键酶。将HSL基因作为影响猪脂肪代谢和沉积相关性状的候选基因,对不同品种猪HSL基因 5′ UTR和外显子Ⅰ片段进行了克隆测序并开展多态性与性状间的关系研究。序列比较发现,在测定的HSL基因靠近起始密码子(ATG)的 419bp中,杜洛克、梅山猪、大白猪和清平猪序列完全一致,与长白猪序列比较,在-13 ^-12bp位置存在GC→CG的碱基变异;梅山猪(3个体)、通城猪(3个体 )、长白猪 (3个体 )、大白猪(3个体)HSL基因外显子Ⅰ的 442bp位置有G→A碱基间的变异,G→A的转换改变了限制性内切酶BsaHⅠ酶识别位点,且导致了编码氨基酸Val→Ile的替换。经PCR RFLP分析,HSL基因外显子ⅠBsaHⅠ位点多态性有AA、AG和GG 3种基因型。“大白×梅山”F2 代资源家系猪BsaHⅠ位点不同基因型个体背膘厚、肌内脂肪等性状协方差统计分析发现,AG基因型和GG基因型在眼肌面积上存在显著差异。

Hormone-sensitive lipase (HSL) is the key enzyme responsible for the mobilization of free acids from adipose tissue,and it is also the most important enzyme that affect fat deposition.In this paper,the porcine hormone-sensitive lipase gene 5′-UTR and exon Ⅰ were sequenced.The sequence number in GenBank are AY332499,AY332497,AY332504,AY332505.A GC→CG in the DNA sequence -13^-12 bp of porcine HSL gene 5′-UTR was detected between Duroc,Meishan,Qingping pig, Largewhite and Landrace.A G→A missense mutation was detected in HSL gene exonⅠ of different pig breeds. The characterization of the BsaHⅠ PCR-RFLP polymorphism in exonⅠ of the porcine HSL gene of different breeds and “Largewhite×Meishan” F_2 group was analyzed.By association analysis between BsaHⅠ PCR-RFLP polymorphism and GG,GG,AA genotypes of HSL gene exonⅠ,a significant difference of pig eye area was found between AG and GG genotypes (P<0.05) in F_2 group.