Cloning and sequencing of the full-length of human PPAR? cDNA
- VernacularTitle:人PPAR?受体cDNA全长序列的克隆及测序
- Author:
Tao ZHANG
- Publication Type:Journal Article
- Keywords:
Peroxisome proliferator-activated receptor ?;
RT-PCR;
cDNA clone;
T vector;
Sequencing
- From:
Journal of Chongqing Medical University
2007;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone and sequence the full-length of human PPAR? cDNA in order to further construct the eukaryotic expression vector carrying hPPAR? gene.Methods:hPPAR? cDNA was cloned from HepG2 cells total RNA by RT-PCR.The PCR product recovered from gel were ligated with pMD19-T vector and transformed into DH5? competent cell.The integrity and fidelity of hPPAR? cDNA sequence inserted in T vector were verified by BamH I and Sal I double excising and DNA sequencing assays.Results:The positive clone T vector plasmid containing correct sequence of hPPAR? cDNA were verified by enzyme digestion as well as sequence analysis and was named as pMD19-hPPAR?-T vector.The sequence of inserted hPPAR? cDNA was in accordance with the corresponding sequence in GeneBank database(AY919140).Conclusion:hPPAR? gene is successfully cloned and the pMD19-hPPAR?-T intermediate vector is constructed,which provide a good basis for further constructing the eukaryotic expression vector carrying hPPAR? gene and studying the receptor's function.
