小鼠干扰素γ基因克隆、测序及辐射诱导表达质粒pIRESEgr-IFNγ的构建

Cloning and sequencing of mouse IFN-γ and construction of radiation-inducible expression plasmid pIRESEgr-IFNγ

目的 :克隆小鼠干扰素γ (IFN -γ)编码区 c DNA序列并构建含 Egr- 1启动子的辐射诱导表达质粒p IRESEgr- IFNγ。方法 :利用逆转录多聚酶链反应 (RT- PCR)法 ,以小鼠脾细胞 m RNA为模板 ,扩增获得全长IFNγc DNA,与 p GEMT载体连接做全自动测序 ,并利用基因重组技术构建含 Egr- 1启动子的辐射诱导表达质粒p IRESEgr- IFNγ。结果 :经测序证实获得的小鼠 IFNγ c DNA序列与文献报道完全一致 ,并构建了含 Egr- 1启动子的辐射诱导表达质粒 p IRESEgr- IFNγ。结论 :成功克隆了小鼠 IFNγ的 c DNA序列 ,构建了辐射诱导表达质粒p IRESEgr-

ObjectiveTo clone the sequence of the cDNA of mouse interferon gamma coding area and construct radiation inducible expression plasmid pIRESEgr IFNγ containing Egr 1 promoter. MethodsWith the technique of RT PCR, mouse splenocyte mRNA was used as template to obtain full length mouse IFNγ. The pGEMT IFNγ was sequenced automatically and radiation inducible expression plasmid pIRESEgr IFNγ containing Egr 1 promoter was constructed with gene recombinant technique. ResultsThe sequencing proved the cloned mouse IFNγ cDNA to be completely identical with that reported in the literature and the radiation inducible expression plasmid pIRESEgr IFNγ containing Egr 1 promoter was constructed successfully. ConclusionMouse IFNγ cDNA was cloned and the radiation inducible expression plasmid pIRESEgr IFNγ containing Egr 1 promoter was constructed successfully.