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植物细胞表达人胰岛素的载体构建
引用本文:汪春义,戚凤春,陈桂玲,申镇维,王宣军,盛军.植物细胞表达人胰岛素的载体构建[J].粉末涂料与涂装,2006,19(4):344-346.
作者姓名:汪春义  戚凤春  陈桂玲  申镇维  王宣军  盛军
作者单位:吉林大学生命科学学院,吉林大学生命科学学院,长春生物制品研究所,长春生物制品研究所,长春生物制品研究所,长春生物制品研究所 长春130021,长春生物制品研究所长春130062,长春130021,长春生物制品研究所长春130062,长春130062,长春130062,长春130062,长春130062
摘    要:目的构建人胰岛素的植物细胞表达载体。方法设计多对引物,通过PCR反应合成霍乱毒素B亚单位(CTB)基因和不带C肽的人胰岛素基因。将霍乱毒素B亚单位(CTB)基因与这种不带C肽的人胰岛素基因融合后,插入克隆载体,然后用BamHⅠ和SacⅠ进行酶切,并将酶切的融合基因片段连接在植物表达载体pBI121上,最后用BamHⅠ和SacⅠ进行酶切鉴定。结果测序结果表明,PCR扩增的目的基因顺序与设计完全一致,并构建了克隆载体和植物表达载体。植物表达载体经酶切后鉴定,所含基因片段大小与预期相符。结论已成功地构建了人胰岛素基因的植物细胞表达载体。

关 键 词:人胰岛素基因  霍乱毒素B亚单位(CTB)  植物表达载体
收稿时间:2006-05-12
修稿时间:2006年5月12日

Construction of Vector for Expression of Human Insulin in Plant Cells
WANG Chun-yi, QI Feng-chun, CHEN Gui-ling, et al.Construction of Vector for Expression of Human Insulin in Plant Cells[J].Chinese Journal of Biologicals,2006,19(4):344-346.
Authors:WANG Chun-yi  QI Feng-chun  CHEN Gui-ling  
Affiliation:College of Life Science, Jilin University, Changchun 130012, China
Abstract:Objective To construct the vector for expression of human insulin in plant cells.Methods The cholera toxin B subunit(CTB) gene and human insulin gene without C peptide were synthesized by PCR using the designed primers,then fused and inserted into cloning vector pMD18-T.Digest the recombinant plasmid with BamHⅠ and SacⅠ,and clone the obtained fusion gene into expression vector pBI121.Identify the recombinant plasmid by digestion with BamHⅠ and SacⅠ.Results The sequences of synthetic CTB and human insulin genes were completely identical to those designed.Restriction map proved that the length of gene fragment inserted into expression vector pBI121 was consistent with that expected.Conclusion The vector for expression of human insulin in plant cells was successfully constructed.
Keywords:Human insulin gene  Cholera toxin B subunit(CTB)  Plant expression vector  
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