山羊SKIP基因的分离、鉴定及与生长性状的关联分析

发布时间：2018-02-21 04:28

本文关键词： 单核苷酸多态性位点山羊骨骼肌和肾脏富集的肌醇磷酸酶生长性状成肌分化　出处：《华中农业大学学报》2017年06期 　论文类型：期刊论文

【摘要】：为研究SKIP基因对山羊生长发育的影响,并寻找与山羊生长性状相关的分子标记,利用绵羊与牛SKIP基因的同源序列设计引物,RT-PCR扩增得到一段长1 350bp的山羊SKIP基因表达序列,包括其完整编码区序列,编码449个氨基酸的蛋白。通过序列Blast比较和系统发育树的构建,发现山羊SKIP与绵羊SKIP序列相似性最高(99.52%)。利用生物信息学分析SKIP蛋白的理化性质,预测SKIP蛋白分子质量为52ku,等电点为5.18,包含典型的疏水区域和磷脂酰肌醇5-磷酸磷酸酶结构域,定位在细胞质(60.9%)和细胞核(26.1%)中。间接免疫荧光试验验证了SKIP的细胞定位,并发现位于细胞核中的SKIP在成肌细胞分化过程中转移到细胞质。通过波尔山羊和麻城黑山羊的DNA混池测序,发现SKIP内含子2有一处G/A突变,利用PCRRFLP方法对波尔×麻城黑山羊F2代杂交后代进行基因分型,性状关联分析发现该单核苷酸多态性位点(SNP)与出生体质量、胸围、管围、身高等性状显著相关,等位基因G与生长呈正相关。
[Abstract]:In order to study the effect of SKIP gene on the growth and development of goats, and to search for molecular markers related to goat growth traits, a 1 350 BP long goat SKIP gene was amplified by RT-PCR using primers designed by homologous sequence of sheep and cattle SKIP gene. By comparing sequence Blast and constructing phylogenetic tree, it was found that goat SKIP and sheep SKIP sequence had the highest similarity (99.52%). The physicochemical properties of SKIP protein were analyzed by bioinformatics. The predicted molecular weight of SKIP protein is 52ku.The isoelectric point is 5.18. it contains a typical hydrophobic region and phosphatidylinositol 5-phosphatase domain, located in cytoplasm 60.9 and nucleus 26.1). Indirect immunofluorescence assay verifies the cellular localization of SKIP. It was found that SKIP located in the nucleus was transferred to the cytoplasm during myoblast differentiation. By sequencing the DNA mixed pool of Boer goat and Macheng black goat, we found a G / A mutation in intron 2 of SKIP. The genetic analysis of F2 progeny of Boer 脳 Macheng Black Goat was carried out by PCRRFLP method. The trait association analysis showed that the single nucleotide polymorphism (SNP) was significantly correlated with birth weight, chest circumference, tube circumference, height and other traits. Allele G was positively correlated with growth.
【作者单位】：湖北省农业科学院畜牧兽医研究所/动物胚胎工程及分子育种湖北省重点实验室;华中农业大学农业动物遗传育种与繁殖教育部重点实验室/农业部猪遗传育种重点开放实验室/生猪健康养殖协同创新中心;
【基金】：国家自然科学基金青年项目(31402051)
【分类号】：S827

【相似文献】