| 真核生物多顺反子表达载体的构建及在Hela细胞中的表达 |
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| 引用本文： | 苗素平,苗素生,扈廷茂,刘明秋.真核生物多顺反子表达载体的构建及在Hela细胞中的表达[J].内蒙古大学学报(自然科学版),2009,40(1). |
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| 作者姓名： | 苗素平 苗素生 扈廷茂 刘明秋 |
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| 作者单位： | 1. 内蒙古大学生命科学学院,呼和浩特,010021
2. 哈尔滨医科大学附属肿瘤医院头颈外科,哈尔滨,150040 |
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| 基金项目： | 内蒙古自治区自然科学基金 |
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| 摘 要： | 利用DNA重组技术将抗肿瘤血管生成的内皮抑素基因endostatin插入含有FHIT(抗肿瘤人脆性组氨酸三联体基因)和EGFP的pIRES2-FHIT-EGFP载体中构建了真核多顺反子表达载体pES-IRES-FHIT-IRES-EGFP,用脂质体法转染Hela细胞后,用G418筛选出稳定转染的细胞.经RT-PCR、RT-PCR southern blot、northern blot和免疫细胞化学分析,结果证明单顺反子和三顺反子在RNA水平都得到表达.在蛋白表达水平上,荧光观察和免疫细胞化学分析结果显示Endostatin、FHIT和EGFP在Hela细胞中均得到表达,为基因药物和肿瘤多基因治疗奠定了基础.
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| 关 键 词： | 多顺反子 内皮折素基因 |
Construction of Eukaryote Polycistronic Expression Vector and Expression in Hela |
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| MIAO Su-ping,MIAO Su-sheng,HU Ting-mao,LIU Ming-qiu.Construction of Eukaryote Polycistronic Expression Vector and Expression in Hela[J].Acta Scientiarum Naturalium Universitatis Neimongol,2009,40(1). |
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| Authors: | MIAO Su-ping MIAO Su-sheng HU Ting-mao LIU Ming-qiu |
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| Institution: | 1.College of Life Sciences;Inner Mongolia University;Hohhot 010021;China;2.Department of Head-Neck Surgery;Third Hospital of Harbin Medical University;Harbin 150040;China |
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| Abstract: | Endostatin cDNA was obtained by RT-PCR and then inserted into pEF1-4SX.Then the fragment of endostatin linking with IRES was cut and ligated into a eukaryotic expression vector to construct the tricistronic eukaryotic expression vector pES-IRES-FHIT-IRES-EGFP.This recombinant vector was transfected into Hela cells with LipofectamineTM 2000.Total RNA was extracted from those stably transfected Hela cells,and then was checked by RT-PCR Southern blotting.The results indicated that the moncistrons of endostatin... |
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| Keywords: | IRES FHIT EGFP |
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