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猪白血病抑制因子的真核表达与活性分析
引用本文:李明堂,付玉,王清爽,果洪宇,李洪广. 猪白血病抑制因子的真核表达与活性分析[J]. 吉林农业大学学报, 2008, 30(6)
作者姓名:李明堂  付玉  王清爽  果洪宇  李洪广
作者单位:长春理工大学生命科技学院,长春,130022;吉林大学中日联谊医院神经外科,长春,130031
基金项目:吉林省科技发展计划项目(20010522);;长春市科技计划项目(2006075)
摘    要:
采用分子克隆技术构建猪白血病抑制因子(LIF)的真核表达载体,表达重组猪LIF蛋白并分析其活性。酶切和DNA测序显示,所构建的真核表达质粒pcDNA3.1-pLIF—MycHis含有编码分泌型猪LIF蛋白的CD—NA序列;免疫印迹分析显示,分泌到上清中的重组蛋白在相对分子质量约37000的位置上有明显的增强表达条带;活性分析结果显示,重组蛋白能够维持小鼠E-14胚胎干细胞在未分化状态。该结果显示重组猪UF蛋白在CHO细胞中进行了正确表达并具有很好的生物活性,能够用来培养小鼠胚胎干细胞,为用于猪胚胎干细胞系的研究奠定了良好的基础。

关 键 词:猪白血病抑制因子  活性分析  胚胎干细胞

Eukaryotic Expression of Porcine LIF and the Assay of Its Bioactivities
LI Ming-tang,FU Yu,WANG Qing-shuang,GUO Hong-yu,LI Hong-guang. Eukaryotic Expression of Porcine LIF and the Assay of Its Bioactivities[J]. Journal of Jilin Agricultural University, 2008, 30(6)
Authors:LI Ming-tang  FU Yu  WANG Qing-shuang  GUO Hong-yu  LI Hong-guang
Affiliation:1.School of Life Science and Technology;Changchun University of Science and Technology;Changchun 130022;China;2.Department of Neurosurgery;China-Japan Union Hospital;Jilin University;Changchun 130031;China
Abstract:
The expression plasmid of porcine LIF in the eukaryotic system was constructed by molecular cloning technique,the recombinant porcine LIF protein was expressed exactly,and the bioactivity of recombinant porcine LIF protein was further assayed immediately.The results indicated that the constructed expression plasmid pcDNA3.1-pLIF-MycHis was identified by DNA sequencing and restriction enzyme digestion;the expression of the recombinant porcine LIF protein was performed exactly in CHO cell;in the conditioned m...
Keywords:LIF  activity analysis  ES cell  
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