| 人肝再生增强因子基因的克隆和真核表达载体的构建 |
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| 引用本文： | 李雪梅,赵春丽.人肝再生增强因子基因的克隆和真核表达载体的构建[J].河北农业大学学报,2005,28(5):85-88. |
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| 作者姓名： | 李雪梅 赵春丽 |
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| 作者单位： | 1. 河北农业大学,动物科技学院,河北,保定,071000;中国科学院,动物研究所生殖生物学国家重点实验室,北京,100080
2. 中国科学院,动物研究所生殖生物学国家重点实验室,北京,100080;东北农业大学,动物医学学院生理生化教研室,黑龙江,哈尔滨,150030 |
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| 基金项目： | 黑龙江省重大科技攻关项目（GB01C106-02）. |
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| 摘 要： | 从6月龄流产的胎儿肝脏中提取总RNA,利用反转录-PCR(RT-PCR)技术扩增出人肝细胞再生增强因子hALR(Human augmenter of liver regeneration)基因片段,克隆于PET28a(+)载体,经进一步PCR及酶切鉴定,确定为此目的片段后进行序列分析,再将目的片段亚克隆于真核表达载体pcDNA3.1(+),用酶切方法鉴定重组子。采用RT-PCR技术成功地获得了hALR基因片段。序列分析表明,克隆的hALR基因与Genbank报道的人ALR核苷酸序列基本一致。完成了真核表达载体的构建,为hALR基因的真核表达奠定了基础。
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| 关 键 词： | 人肝再生增强因子 克隆 序列分析 真核表达载体 |
| 文章编号： | 1000-1573（2005）05-0085-04 |
| 收稿时间： | 2005-05-12 |
| 修稿时间： | 2005-05-12 |
Cloning and construction of eucaryote expression vector of humanaugmenter of liver regeneration gene |
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| LI Xue-mei,ZHAO Chun-li.Cloning and construction of eucaryote expression vector of humanaugmenter of liver regeneration gene[J].Journal of Agricultural University of Hebei,2005,28(5):85-88. |
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| Authors: | LI Xue-mei ZHAO Chun-li |
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| Institution: | 1. College of Animal Science, Agricultural University of Hebei, Baoding 071001, China; 2. State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Science, Beijing 100080, China; 3. Biochemical and Physiological Division of Animal Medicine Sciences College, Northeast Agricultural University, Harbin 150030, China |
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| Abstract: | To clone the fragment of human liver regeneration(hALR) gene and construct the eucaryote expression vector for hALR expression,we extract total RNA from 6-month old fetal liver.and amplified hALR gene fragment by RT-PCR technique,The PCR product was cloned into PET28a(+) vector and recombinants were identified by PCR and enzyme digestion.The analyzed gene sequence showed that it has the basic sequence with the one in Genebank.The target gene was subcloned into eucaryote expression vector pcDNA3.1(+) and recombinants were identified by enzyme digestion.The successfully construction of the eucaryote expression vector provide the foundation of hALR eucaryote expression. |
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| Keywords: | human augmenter of liver regeneration clone sequence analysis eucaryote expression vector |
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