鹅LPL基因外显子4、5、6克隆及序列分析

Cloning and analyzing exon 4,5 and 6 of lipoprotein lipase gene in goose

为揭示鹅脂蛋白脂酶（LPL）脂质和肝素结合区域的特性,对鹅LPL基因外显子4-6进行了克隆和序列分析。以皖西白鹅血清总DNA为模板,设计引物对鹅LPL基因的第4-6外显子区域进行扩增和测序,分析鹅与其他物种之间该区域的DNA序列同源性、密码子非随机应用以及相应氨基酸残基区域的亲水性。结果表明,鹅LPL基因外显子4-6的片段大小分别为112、234和243 bp,其序列与鸡LPL基因相应序列的同源性达到91.9%,与人、羊、牛、猪等哺乳动物的同源性为74%-77%;鹅与鸡密码子非随机应用的相似系数为0.839,与猪和鼠的相似系数为0.580-0.650;LPL中由外显子4-6编码的氨基酸残基有4个区域具有较大的表面概率,分别位于残基145-148、残基187-190、残基255-257和残基293-299区域,表面概率分别1.62-4.29、2.06-3.01、2.75-4.46和3.03-6.01,这些区域伴随有较强的亲水性。

Exon 4 to intron 6 sequence of lipoprotein lipase （LPL） gene was cloned to determine genome characteristics in goose. The total DNA from goose serum was extracted. Two pairs of primers, P and Q, were designed to amplify the LPL gene exon 4 to intron 6 which were then cloned and sequenced. The results showed that the LPL gene exon 4 to intron 6 were 3 404 bp. Blasting the sequence homology among chicken, human, pig, mouse, cattle and cat showed that the DNA homology between goose and chicken was 91.9% which was the highest between goose and other species. The homology between goose and mammal （human, sheep, cattle and pig） was 74 to 77%. Also the hydrophilicity, average charge, surface probability and condon usage of amino acides coded by LPL gene exon 4 to exon 6 in goose were analysed.