人Flt3全长基因的克隆及其逆转录病毒表达载体的构建  

Cloning of Human Flt3 Full-length Gene and Construction of Its Retroviral Expressing Vector

作　　者：居颂光[1] 居颂文[1] 仇红霞[1] 傅晋祥[1] 周斌[1] 王凤鸣[1] 朱华亭[1] 胡玉敏[1] 刘彤[1] 刘高勤[1] 张学光[1] 

机构地区：[1]苏州大学医学生物技术研究所,江苏苏州215007

出　　处：《中国血液流变学杂志》2005年第4期544-547,578,共5页Chinese Journal of Hemorheology

基　　金：国家自然基金青年科学基金项目(30400394);江苏省放射医学与防护重点实验室开放经费资助项目(KJS05031);苏州大学医学发展基金资助项目(EE132031)

摘　　要：目的克隆人Flt3全长基因,构建PGE/Plt3逆转录病毒表达载体。方法以人骨髓cDNA等基因文库为模板,采用分段克隆、人工PCR延伸及PCR连接等方法,获得了人Flt3全长基因,并构建Flt3基因的逆转录病毒表达载体。结果成功克隆Flt3全长基因和构建PGEZ/Flt3逆转录病毒表达载体。结论F1t3全长基因克隆及构建逆转录病毒表达载体的成功,为构建Flt3转基因细胞和制备抗人Flt3单克隆抗体和研究Flt3的生物学功能奠定了物质基础,也为低丰度和长片段基因克隆提供了有价值的经验。Objective To clone the Flt3 full-length gene and construct its retroviral expressing vector.Methods Overlapped cDNA fragments of Flt3 gene were cloned from human bone marrow cDNA library,extend its 5＇-region step by step,and then linked them together as Flt3 full-length gene by PCR. Full-length Flt3 gene were subcloned to Pstl and BamHl restriction sites of retroviral expressing vector pGEZ-term.Results Cloned the Flt3 full-length gene and construct its retroviral expressing vector PGEZ/Flt3 successfully.Conclusion Flt3 gene was cloned and its expressing vector was constructed successfully.lt provided important value to establish Flt3 engineering cell lines,prepare anti-human Flt3 monoclonal antibody and study its biological functions.

关 键 词：FLT3 CD135 基因 克隆 

分 类 号：R392.4[医药卫生—免疫学]