从江香猪ApoC3基因的克隆及亚细胞定位研究

发布时间：2018-02-04 03:18

本文关键词： 从江香猪载脂蛋白C基因基因克隆亚细胞定位　出处：《畜牧与兽医》2017年08期 　论文类型：期刊论文

【摘要】：为了观察从江香猪载脂蛋白C3(ApoC3)基因及其表达产物在真核细胞中的亚细胞定位情况,以脂肪型小型猪从江香猪为研究对象,克隆ApoC3基因CDS区,构建携带有绿色荧光蛋白的pEGFP-C1-ApoC3重组质粒,转染HEK-293T细胞。结果显示:从江香猪ApoC3基因与GenBank上公布的序列相比,在141位有1个碱基发生转换,由腺嘌呤(A)突变成鸟嘌呤(G),为无义突变;对ApoC3蛋白的亚细胞定位分析表明,该蛋白在胞外基质占55.6%,内质网占22.2%,液泡占11.1%,细胞质占11.1%。pEGFP-C1-ApoC3重组质粒转染HEK-293T细胞后的荧光共定位结果显示,ApoC3蛋白在细胞中表达部位与PSORT II Prediction软件预测结果一致。本研究结果为进一步构建ApoC3基因转基因动物模型,开展ApoC3基因的多态性变化而导致的相关疾病研究奠定了基础。
[Abstract]:In order to observe the subcellular localization of apolipoprotein C _ 3 (ApoC _ 3) gene and its expression products in eukaryotic cells of Congjiang Xiang pig, the fatty miniature pig Congjiang Xiang pig was studied. The CDS region of ApoC3 gene was cloned and the recombinant plasmid of pEGFP-C1-ApoC3 carrying green fluorescent protein was constructed. The results of transfection of HEK-293T cells showed that there was one base conversion between the ApoC3 gene of Congangxiang pig and the published sequence on GenBank. From adenine (A) to guanine guanine (G), it is a nonsense mutation. The subcellular localization analysis of ApoC3 protein showed that the protein was 55.6% in extracellular matrix, 22.2% in endoplasmic reticulum and 11.1% in vacuole. The fluorescence co-localization of the recombinant plasmid pEGFP-C1-ApoC3 after transfection into HEK-293T cells showed that cytoplasm accounted for 11.1%. The expression site of ApoC3 protein in cells was consistent with that predicted by PSORT II Prediction software. This study was intended to further construct the transgenic animal model of ApoC3 gene. The research on the related diseases caused by the polymorphism of ApoC3 gene has laid a foundation.
【作者单位】：高原山地动物遗传育种与繁殖省部共建教育部重点实验室;贵州大学动物科学学院;贵州露元现代农业发展有限公司;
【基金】：省校联合基金(黔科合LH字[2014]7668) 贵州大学青年基金(贵大自青基合字[2012]010号)
【分类号】：Q78;S828
【正文快照】： 脂蛋白主要由胆固醇、甘油三酯(TG)、磷脂和载脂蛋白组成。载脂蛋白是指位于脂蛋白中的蛋白质,目前已发现20余种。载脂蛋白能介导脂蛋白与细胞膜上的脂蛋白受体结合,并被摄入细胞内进行分解代谢[1-2]。载脂蛋白C3(Apo C3)是脂蛋白代谢的重要调节剂,被认为是血脂代谢中与脂蛋白

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