人类p73基因TAp73αcDNA片段克隆及鉴定

Cloning and Identification of TAp73αcDNA Fragment

目的 构建真核表达重组质粒pcDNA3 1+ /TAp73αcDNA ,克隆包含人类 p73基因TAp73α转录本蛋白编码区的cDNA片段。方法 采用RT PCR方法获取TAp73αcDNA目的片段 ,构建pcDNA3 1+ /TAp73αcDNA重组质粒 ,用构建的重组质粒转化大肠杆菌JM 10 9,通过酶切和测序进行相应的鉴定。 结果 包含人类 p73基因TAp73α转录本蛋白编码区的cDNA片段被成功插入真核表达载体 pcDNA3 1+ 质粒的多克隆位点 ,经鉴定与理论设计完全一致。结论 包含人类 p73基因TAp73α转录本蛋白编码区的cDNA片段已被成功克隆。

Objective To clone of the p73 gene cDNA fragment which encodes TAp73α protein.Methods Total RNA were isolated from human SH-SY5Y cell. TAp73α cDNA fragment was amplified by reverse transcription-polymerase chain reaction(RT-PCR), and was inserted into eukaryotic expression vector pcDNA3.1+, and then the recombinant pcDNA3 1+/ TAp73α cDNA was transformed into E.coli JM109 host bacteria.Results The TAp73α cDNA fragment amplified by RT-PCR was proved to be consistant with that reported in Genbank [NM005427]. Restriction enzyme digestion analysis show that the recombinant pcDNA3 1+/ TAp73α cDNA was successfully transformed into E.coli JM109 host bacteria.Conclusion p73 gene cDNA fragment which encodes TAp73α protein was inserted into eukaryotic expression vector pcDNA3 1+ and was successfully cloned.