Cloning and expression of xeroderma pigmentosum complementation group A cDNA and the identification of its recombinant protein.
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.


CONCLUSIONS
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.
Yang
Yong
Y
Department of Dermatology, Peking University First Hospital, Beijing 100034, China. yongyang81@yahoo.com.cn
Bu
Dingfang
D
Wang
Ke
K
Tu
Ping
P
Zhu
Xuejun
X
D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli; D005822:Genetic Vectors; D006801:Humans; D011993:Recombinant Fusion Proteins; D051760:Xeroderma Pigmentosum Group A Protein / Q000235:genetics*
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein