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Updated: Aug 14, 2026
Subcloning Plus Insertion (SPI) - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors
Published on: January 8, 2015
Cloning and expression of xeroderma pigmentosum complementation group A cDNA and the identification of its
Yong Yang1, Dingfang Bu, Ke Wang
1Department of Dermatology, Peking University First Hospital, Beijing 100034, China. yongyang81@yahoo.com.cn
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.
