| 鲫鱼一种新型SNRPC基因的蛋白表达和抗体制备分析 |
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| 引用本文： | 王红莹,杜嫚.鲫鱼一种新型SNRPC基因的蛋白表达和抗体制备分析[J].河北农业科学,2010,14(12):43-45. |
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| 作者姓名： | 王红莹 杜嫚 |
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| 作者单位： | 中南民族大学生命科学学院,湖北武汉430074 |
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| 摘 要： | 本研究构建了鲫鱼卵母细胞特异表达的新型SNRPC基因的原核表达载体,表达并纯化蛋白,制备多克隆抗体,并检测抗体特异性,为进一步研究新型SNRPC基因的功能奠定基础。首先设计引物,应用RT-PCR从鲫鱼成熟卵母细胞中获得含该基因片段,重组入原核表达载体PinPoint T中,在大肠杆菌BL-21(DE3)中诱导表达,应用亲和层析法获得纯度较高的原核表达蛋白并免疫家兔制备多克隆抗体,用Western blot方法检测此抗体特异性。成功构建了新型SNRPC基因重组表达载体,表达的蛋白经纯化后免疫家兔得到了特异的多克隆抗体。鲫鱼卵母细胞的新型SNRPC基因的原核表达载体的构建、重组蛋白的表达纯化及抗体的制备为后续的研究提供了理论基础。
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| 关 键 词： | 鲫鱼 新型SNRPC 蛋白表达 多克隆抗体 抗体 制备 |
A New Type SNRPC Protein Expression and Preparation of Antibody in Carassius carassius gibelio |
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| WANG Hong-ying,DU Man.A New Type SNRPC Protein Expression and Preparation of Antibody in Carassius carassius gibelio[J].Journal of Hebei Agricultural Sciences,2010,14(12):43-45. |
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| Authors: | WANG Hong-ying DU Man |
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| Institution: | Man(College of Life Science,South-central University for Nationalities,Wuhan 430074,China) |
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| Abstract: | The prokaryotic expression vector of differentially expressed gene SNRPC in Carassius carassius was constructed.The recombinant protein was purified to prepare polyclonal antibody.A gene fragment was amplified from cDNA of mature oocyte of crucian carp,inserted into PinPoint T vector and introduced into BL-21（DE3） strain.The recombinant protein was purified by affinity chromatography and then generated the polyclonal antibody with identification by Western blot method.The specific polyclonal antibody was obtained,and it would be used to the functional assay. |
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| Keywords: | Carp New type SNRPC Protein expression Polyclonal antibody Antibody Preparation |
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