Cloning and prokaryotic expression of the cyclophilin gene from Pinus massoniana.

Cyclophilins(CyPs) possess peptidylprolyl cis-trans isomerase activity,which can catalyse the process of protein folding and function as chaperones.CyPs are also involved in other correlated biological such as mRNA procession and signal transduction.A full length cDNA of a cyclophilin gene was cloned from the first strand of Pinus massoniana cDNA by RT-PCR and RACE-PCR methods,named as cyp-pm1(GenBank accession number: GQ497815).The length of cyp-pm1 is 1 002 bp in which the open reading frame has 519 bp nucleotides which encode a deduced protein with 172 amino acid residues,and which has 106 bp and 377 bp untranslated region of 5′end and 3′end respectively,with 26 bp Poly(A) included in 3′ end.The bioinformatic analysis indicated that the pI and MW of animo acids encoded by cyp-pm1 were predicted to be 8.82 and 18 212 respectively,and the protein had one peptidylprolyl cis-trans isomerase binding domain.Cyp-pm1 displayed high sequence identity with previously cloned CyPs family genes of plants.The prokaryotic expression vector of cyp-pm1 gene encoding protein was constructed by subcloning the fragment into pET-24a(+) and was expressed in Escherichia coli induced by IPTG.The molecular weight of the induced protein was about 1.8×104 approximately as predicted.