山羊SKIP基因的分離、鑒定及與生長性狀的關(guān)聯(lián)分析
本文關(guān)鍵詞: 單核苷酸多態(tài)性位點(diǎn) 山羊 骨骼肌和腎臟富集的肌醇磷酸酶 生長性狀 成肌分化 出處:《華中農(nóng)業(yè)大學(xué)學(xué)報》2017年06期 論文類型:期刊論文
【摘要】:為研究SKIP基因?qū)ι窖蛏L發(fā)育的影響,并尋找與山羊生長性狀相關(guān)的分子標(biāo)記,利用綿羊與牛SKIP基因的同源序列設(shè)計引物,RT-PCR擴(kuò)增得到一段長1 350bp的山羊SKIP基因表達(dá)序列,包括其完整編碼區(qū)序列,編碼449個氨基酸的蛋白。通過序列Blast比較和系統(tǒng)發(fā)育樹的構(gòu)建,發(fā)現(xiàn)山羊SKIP與綿羊SKIP序列相似性最高(99.52%)。利用生物信息學(xué)分析SKIP蛋白的理化性質(zhì),預(yù)測SKIP蛋白分子質(zhì)量為52ku,等電點(diǎn)為5.18,包含典型的疏水區(qū)域和磷脂酰肌醇5-磷酸磷酸酶結(jié)構(gòu)域,定位在細(xì)胞質(zhì)(60.9%)和細(xì)胞核(26.1%)中。間接免疫熒光試驗驗證了SKIP的細(xì)胞定位,并發(fā)現(xiàn)位于細(xì)胞核中的SKIP在成肌細(xì)胞分化過程中轉(zhuǎn)移到細(xì)胞質(zhì)。通過波爾山羊和麻城黑山羊的DNA混池測序,發(fā)現(xiàn)SKIP內(nèi)含子2有一處G/A突變,利用PCRRFLP方法對波爾×麻城黑山羊F2代雜交后代進(jìn)行基因分型,性狀關(guān)聯(lián)分析發(fā)現(xiàn)該單核苷酸多態(tài)性位點(diǎn)(SNP)與出生體質(zhì)量、胸圍、管圍、身高等性狀顯著相關(guān),等位基因G與生長呈正相關(guān)。
[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.
【作者單位】: 湖北省農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所/動物胚胎工程及分子育種湖北省重點(diǎn)實(shí)驗室;華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)動物遺傳育種與繁殖教育部重點(diǎn)實(shí)驗室/農(nóng)業(yè)部豬遺傳育種重點(diǎn)開放實(shí)驗室/生豬健康養(yǎng)殖協(xié)同創(chuàng)新中心;
【基金】:國家自然科學(xué)基金青年項目(31402051)
【分類號】:S827
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