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从江香猪ApoC3基因的克隆及亚细胞定位研究

作  者:
卢贤君;赵佳福;苟维胜;段志强;赵焕平
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;贵州露元现代农业发展有限公司;贵州大学动物科学学院
关键词:
从江香猪;载脂蛋白C3基因;基因克隆;亚细胞定位
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。
译  名:
Cloning and subcellular localization of ApoC3 in Congjiang Xiang pigs
作  者:
LU Xianjun;ZHAO Jiafu;GOU Weisheng;DUAN Zhiqiang;ZHAO Huanping;Key Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau Mountainous Region Ministry of Education;College of Animal Science,Guizhou University;Guizhou Lou Yuan Modern Agriculture Development Ltd.Co;
关键词:
Congjiang Xiang pig;;ApoC3 gene;;gene clone;;subcellular localization
摘  要:
In order to investigate sub-cellular localization of ApoC3 gene in eukaryocyte,the fat minipig of Congjiang Xiang pigs were used as a research object and constructed the recombination plasmid pEGFP-C1-ApoC3 and transfected HEK-293T cells. The result showed that one base mutation in 141 bases compared with the gene submission in GenBank,causing adenine( A) to guanine( G),and nonsense mutation was observed. The subcellular localization results of ApoC3 proteins with PSORT Ⅱ prediction showed that the protein expression in the extracellular matrix were 55. 6%,endoplasmic reticulum 22. 2%,vacuole accounted 11. 1% and cytoplasm 11. 1%. The corresponding results were founded by transfected HEK-293T cells with the recombination plasmid pEGFP-C1-ApoC3. Our results will provide a foundation for construction ApoC3 gene transgenic animal models and study of the relationship between ApoC3 gene polymorphisms and related diseases.

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