Tet-on调控的人HOXB4慢病毒载体构建  

The construction of Tet-on regulating lentiviral vector Containing Human Gene HOXB4

作　　者：李秀英[1,2] 白金萍[1] 仲苓芝[1] 李新娜[1] 李文雪[1] 李荣贵[1] 

机构地区：[1]吉林大学基础医学院病理学教研室,吉林长春130021 [2]吉林大学中日联谊医院中心实验室,吉林长春130033

出　　处：《中国实验诊断学》2014年第2期175-178,共4页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金资助项目(21277057)

摘　　要：目的构建Tet-on调控的人HOXB4慢病毒载体,为体外扩增造血干细胞并保持其干细胞特性提供实验材料基础。方法应用PCR技术从质粒TAT-HA-HOXB4-Full中扩增编码人HOXB4的cDNA,用限制性内切酶EcoRI分别酶切HOXB4基因片段和带有Tet-on开关的可调控慢病毒载体,经T4DNA连接酶连接获得慢病毒载体Teto-Fuw-hHOXB4。对构建的HOXB4慢病毒载体进行酶切片段凝胶电泳及DNA测序分析。结果酶切片段电泳分析和DNA测序分析证明Tet-on调控的人HOXB4慢病毒载体构建成功。结论正确构建了Tet-on调控的人HOXB4慢病毒载体,为后续的体外扩增造血干细胞及相关研究提供材料。Objective The recombined lentiviral vector which containing Tet-on and Human HOXB4 was construc- ted in order to provide the material basis for proliferating of hematopoietic stem cells and maintaining properties of stem cells in vitro. Methods Polymerase chain reaction （PCR） was used to amplify HOXB4 gene from the plasmid TAT- HA-HOXB4-Full. Both HOXB4 gene and the lentiviral vector containing Tet-on were digested by EcoRI,recovered,and ligated through T4 DNA ligase to obtain Teto-Fuw-hHOXB4. The recombinant lentiviral vector was checked by restric- tion enzyme digestion and DNA sequencing. Results Both restriction enzyme digestion and DNA sequencing demon- strated that the HOXB4 gene was inserted into the lentiviral vector correctly. Conclusion The Teto-Fuw-hHOXB4 lentiviral vector is constructed,and provided the material basis for proliferating of hematopoietic stem cells in vitro.

关 键 词：HOXB4 慢病毒载体 Tet-on 

分 类 号：R346[医药卫生—基础医学]