人地中海贫血基因β654的克隆与序列分析

Cloning and Sequencing of the Human Thalassemic Gene β654

【目的】克隆人地中海贫血基因 β6 5 4,并进行序列分析 ,为下游转基因小鼠模型的建立奠定工作基础。【方法】以PCR法扩增获得人 β6 5 4基因 ,将其克隆入卸除了人 β基因的质粒 pBGT5 1中 ,酶切、反向点杂交及测序法鉴定重组质粒。【结果】所构建的重组质粒中含人 β6 5 4基因 ,其引入方向正确 ,序列分析结果正确。

In order to establish the foundation for transgenic mouse model, the human thalassemic gene(β654) was cloned and sequenced. The human β654 gene was amplified by PCR, and cloned into the plasmid BGT51 in which the human β gene was cut out aforehand. The recombinant plasmid was certified by enzyme digestion, reverse dot hybridization and sequencing. A recombinant plasmid was obtained, which contained the human β654 gene in the correct recombinant direction. Sequencing showed that the cloned insert was correct. [Conclusions] The recombinant plasmid constructed is useful for establishing a transgenic mouse.