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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 full-length human Flt3 gene and construct PGE / Plt3 retroviral vector. Methods Human full-length cDNA of human Flt3 gene was obtained by gene fragment library of human bone marrow cDNA as a template. The full-length human Flt3 gene was cloned by polymerase chain reaction (PCR) and amplified by PCR. The recombinant retroviral vector containing Flt3 gene was constructed. Results The full-length Flt3 gene was successfully cloned and the PGEZ / Flt3 retrovirus expression vector was constructed. Conclusion The successful cloning of full-length F1t3 gene and construction of retroviral expression vector laid the material foundation for constructing Flt3 transgenic cells and preparing anti-human Flt3 monoclonal antibody and studying the biological function of Flt3, as well as low abundance and long fragment gene Cloning provides valuable experience.