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高等学校化学研究 ›› 2006, Vol. 22 ›› Issue (2): 145-149.

• Articles • 上一篇    下一篇

cDNA Cloning, Prokaryotic and Eukaryotic Expression and Characterization of Porcine Leukemia Inhibitory Factor

LI Ming-tang1, JIANG Yong1, WANG Shu-mei2, LI Yong-ming1, WANG Fang1, HOU Xia1, YANG Hong1, MA Tong-hui1   

  1. 1. Membrane Channel Research Laboratory, Northeast Normal University, Changchun 130024, P.R.China;
    2. Department of Anesthesia, Second Teaching Hospital, Jilin University, Changchun 130041, P.R.China
  • 收稿日期:2005-12-31 出版日期:2006-04-24 发布日期:2011-08-06
  • 基金资助:

    Supported by the National Natural Science Fund for Distinguished Young Scholars(No.30325011), the National Natural Science Foundation of China (Nos.30470405 and 30570864), Distinguished Young Scholars Fund of Jilin Province (No.20030112) and Excellent Young Teachers Program of MOE, P.R.China.

cDNA Cloning, Prokaryotic and Eukaryotic Expression and Characterization of Porcine Leukemia Inhibitory Factor

LI Ming-tang1, JIANG Yong1, WANG Shu-mei2, LI Yong-ming1, WANG Fang1, HOU Xia1, YANG Hong1, MA Tong-hui1   

  1. 1. Membrane Channel Research Laboratory, Northeast Normal University, Changchun 130024, P.R.China;
    2. Department of Anesthesia, Second Teaching Hospital, Jilin University, Changchun 130041, P.R.China
  • Received:2005-12-31 Online:2006-04-24 Published:2011-08-06
  • Supported by:

    Supported by the National Natural Science Fund for Distinguished Young Scholars(No.30325011), the National Natural Science Foundation of China (Nos.30470405 and 30570864), Distinguished Young Scholars Fund of Jilin Province (No.20030112) and Excellent Young Teachers Program of MOE, P.R.China.

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摘要： Molecular cloning of the porcine leukemia inhibitor factor(pLIF) has not been reported.A full-length cDNA encoding pLIF was cloned, expressed and characterized.The full-length porcine LIF cDNA encodes a 202 amino acid protein that has an 84% sequence identity to mouse LIF and 86% sequence identity to human LIF.The deduced amino acid sequence of a pLIF protein contains six conserved consensus N-linked glycosylation sites and six cysteine groups to form potential disulfide bonds.The pLIF was expressed in E coli, as a mature form, and in CHO cells as a secreted form.Both the forms of the recombinant pLIFs can maintain murine embryonic stem cells in an undifferentiated state in a culture.The recombinant pLIFs will be useful in establishing a long-term culture of stable pluripotent porcine embryonic stem cells for further manipulation.

关键词: Leukemia inhibitor factor, Porcine, Cloning, Recombinant protein, Embryonic stem cell, Pluripotency

Abstract: Molecular cloning of the porcine leukemia inhibitor factor(pLIF) has not been reported.A full-length cDNA encoding pLIF was cloned, expressed and characterized.The full-length porcine LIF cDNA encodes a 202 amino acid protein that has an 84% sequence identity to mouse LIF and 86% sequence identity to human LIF.The deduced amino acid sequence of a pLIF protein contains six conserved consensus N-linked glycosylation sites and six cysteine groups to form potential disulfide bonds.The pLIF was expressed in E coli, as a mature form, and in CHO cells as a secreted form.Both the forms of the recombinant pLIFs can maintain murine embryonic stem cells in an undifferentiated state in a culture.The recombinant pLIFs will be useful in establishing a long-term culture of stable pluripotent porcine embryonic stem cells for further manipulation.

Key words: Leukemia inhibitor factor, Porcine, Cloning, Recombinant protein, Embryonic stem cell, Pluripotency