| 人LIF基因的克隆及其真核表达载体的构建 |
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| 引用本文： | 李芳菲,刘永刚,彭彦,王建伟,王亚平. 人LIF基因的克隆及其真核表达载体的构建[J]. 重庆医科大学学报, 2005, 30(1): 102-103,107 |
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| 作者姓名： | 李芳菲 刘永刚 彭彦 王建伟 王亚平 |
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| 作者单位： | 重庆医科大学基础医学院,重庆市生物化学与分子药理学重点实验室,重庆,400016;重庆医科大学基础医学院,重庆市生物化学与分子药理学重点实验室,重庆,400016;重庆医科大学基础医学院,重庆市生物化学与分子药理学重点实验室,重庆,400016;重庆医科大学基础医学院,重庆市生物化学与分子药理学重点实验室,重庆,400016;重庆医科大学基础医学院,重庆市生物化学与分子药理学重点实验室,重庆,400016 |
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| 基金项目： | 重庆医科大学校科研和教改项目 |
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| 摘 要： | 目的:克隆白血病抑制因子(LIF)基因,测定其序列,构建带LIF基因的真核表达载体.方法:通过逆转录-聚合酶链式反应(RT-PCR)方法从正常妊娠的子宫蜕膜组织中克隆LIF基因,与T载体连接,再酶切T-LIF载体,获得LIF基因,然后将其与质粒PCDNA3.1( )连接,构建PCDNA3.1( )-LIF真核表达载体.结果:经酶切和DNA测序证明LIF基因序列正确,载体构建成功.结论:成功克隆了LIF基因,构建了PCDNA3.1( )-LIF真核表达载体,为LIF在相应真核细胞内表达及LIF蛋白的分离纯化奠定了基础.
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| 关 键 词： | 白血病抑制因子 RT-PCR 基因克隆 真核表达 |
| 文章编号： | 0253-3626(2005)01-0102-02 |
The clone of human LIF gene and construction of its eukaryotic expression vector |
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| LI Fangfei,et al. The clone of human LIF gene and construction of its eukaryotic expression vector[J]. Journal of Chongqing Medical University, 2005, 30(1): 102-103,107 |
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| Authors: | LI Fangfei et al |
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| Abstract: | Objective:To clone LIF gene for the construction of mammalian expression vector PCDNA3.1(+)-LIF.Methods:LIF gene was cloned from pregnant uterine decidua tissues by using RT-PCR method,the gene was ligated with T-vector,then LIF gene was excised from T-LIF plasmid and inserted into PCDNA3.1(+) to construct eukaryotic expression vector consisting of LIF gene.Results:LIF gene sequence and vector were verified correctly by enzyme digestion,sequence analysis.Conclusion:The successful clone of LIF gene and the construction of its mammalian expression vector has been obtained.The results have provided basis for the further studies on LIF. |
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| Keywords: | LIF RT-PCR Gene clone Mammalian expression |
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