Abstract:

Objective To clone cholesteryl ester transfer protein (CETP) gene of rabbits and to construct its eukaryotic expression vector,and to study the st
ructure and function of CETP gene,and to lay the foundation for further study on the action mechanism of CETP in atherosclerosis(AS).
Methods Total RNA of rabbit liver tissue  was extracted by  TRIzol.RNA was  reversely transcripted into  cDNA by RT-PCR method.Then the CETP gene was cloned by PCR amplification with LA Taq enzyme and the cDNA was regared as template.The CETP cDNA fragment was
cloned into the pMD19-T vector.The recombinant CETP-pMD19-T was sequenced and alignmented with GenBank sequence.The sec
ondary structure of CETP was analyzed by nnPredict software.Then the CETP cDNA fragment was cloned into the pcDNA3.1 eukaryotic expression vector.And the recombinant plasmid was identified by restriction enzyme and PCR.Results The  sequencing and matching  results showed that  it w
as 98%  corresponded to M27486 gene.The full length of CETP cDNA was 1 659 bp,and the length of coded sequence(CDs zone) was 1
 494 bp,and it encoded 498 amino acids. The secondary structure of CETP was the α-helix structure mainly,its molecular weight was 142 119.4，and pI was 4.91.Conclusion The CETP cDNA is successfully cloned，and the CETP eukaryotic expression vector is constructed and the secondary structure of CETP is forecasted  successfully.

Key words: cholesteryl ester transfer protein, cloning, sequence analysis, eukaryotic expression vector