摘要
目的 :克隆、表达A组着色性干皮病基因 ,鉴定重组蛋白。方法 :以人扁桃腺的cDNA为模板 ,RT -PCR及巢式PCR扩增A组着色性干皮病基因的全部编码区 ,克隆入pBluescript质粒 ,DNA测序鉴定。酶切并连接到pTrcHisC载体 ,在大肠杆菌TOP10中表达。免疫印迹鉴定表达产物。 结果 :克隆了A组着色性干皮病基因的完整编码序列 ,表达了预期分子量的融合蛋白 ,经免疫印迹法检测具有免疫原性。结论 :成功克隆了A组着色性干皮病基因 ,表达了其编码蛋白。
Objective: To clone and express xeroderma pigmentosum, complementation group A ( XPA ) cDNA and to identify its recombinant protein. Methods: Coding region of XPA cDNA was amplified from human tonsil cDNA by reverse transcription and nested PCR. The PCR product was cloned into pBluescript vector, confirmed by DNA sequencing, cloned in frame into a prokaryotic expression vector pTrcHis C and expressed in E. coli TOP10. The recombinant fusion protein was identified by immunoblotting. Results: The entire coding region of XPA cDNA was cloned and expressed. The fusion XPA protein was identified by anti XPA monoclonal antibody on western blot. Conclusion: Cloning of human XPA cDNA and successful expression of recombinant XPA protein will be useful for the construction of a viral gene transfer vector for the gene therapy of XPA.
出处
《北京大学学报（医学版）》
CAS
CSCD
北大核心
2003年第4期426-428,共3页
Journal of Peking University:Health Sciences
基金
SupportedbySpecialfundforpromotionofeducation
MinistryofEducationP .R .C
关键词
A组着色性干皮病基因
克隆
表达
重组蛋白
鉴定
Xeroderma pigmentosum
Cloning,molecular
Gene expression
Recombinant proteins/isol
