| 弓形虫AMA1蛋白真核表达载体的构建及免疫活性检测 |
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| 引用本文： | 金春梅,张厚双,薛书江,权根花,崔金伟,王兴贵.弓形虫AMA1蛋白真核表达载体的构建及免疫活性检测[J].安徽农业科学,2010,38(29):16243-16244,16255. |
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| 作者姓名： | 金春梅 张厚双 薛书江 权根花 崔金伟 王兴贵 |
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| 作者单位： | 延边大学农学院动物医学系,吉林延吉,133000;中国农业科学院上海兽医研究所动物寄生虫病研究室,上海,200241;辽宁省动物卫生监督所,辽宁沈阳,110169 |
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| 摘 要： | 目的]构建弓形虫AMA1基因的真核表达质粒,研究其在体外细胞中的表达情况。方法]运用PCR方法和克隆技术,构建pcD-NA3.1-AMA1重组质粒;利用脂质体介导转染法,将该重组质粒导入HEK-293细胞,用Western blotting法检测其在体外的表达情况。结果]从弓形虫RH株cDNA中扩增出AMA1基因片段,成功构建重组表达质粒pcDNA3.1-AMA1,转染AMA1基因的HEK-293细胞表达产物经Western blotting鉴定,其分子量约为70kD,能被特异性免疫血清所识别。结论]构建的真核表达质粒pcDNA3.1-AMA1能在HEK-293细胞中表达,为下一步弓形虫DNA疫苗的研究奠定了基础。
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| 关 键 词： | 弓形虫 AMA1基因 真核表达 |
Eukaryocyte Expression Vector Construction and Immunocompetence Test of AMA1 Protein in Toxoplasma |
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| JIN Chun-mei et al.Eukaryocyte Expression Vector Construction and Immunocompetence Test of AMA1 Protein in Toxoplasma[J].Journal of Anhui Agricultural Sciences,2010,38(29):16243-16244,16255. |
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| Authors: | JIN Chun-mei |
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| Institution: | JIN Chun-mei et al(Department of Veterinary Medicine,Agricultural College,Yanbian University,Yanji,Jilin 133000) |
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| Abstract: | Objective]To construct the recombinant eukaryotic expression plasmid containing the Toxoplasma gondii apical membrane antigen 1(TgAMA1)and study the expression of this recombinant plasmid in HEK-293.Method]The specific fragment of AMA1 gene from the cDNA of T.gondii RH strain was amplified by PCR,and then the amplified fragment was subcloned into pcDNA3.1 expression vector.The eukaryotic expression plasmid pcDNA3.1-AMA1 was then transferred into HEK-293 cells by liposome method.pcDNA3.1-AMA1 expressed pro... |
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| Keywords: | Toxoplasma AMA1 gene Eukaryotic expression |
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