斑节对虾亲环素A(cyclophilinA)基因的克隆及启动子序列分析

CyclophilinsA is a family of proteins that bind the immunosuppressive agent cyclosporin A (CsA) with high affinity and belong to the superfamilies of peptidyl-prolyl cis-trans isomerases (PPIase). CypA has highly conserved structures and exhibits various biological activities which have an important role in cell cycle events. The PPIase activity of CypA can bind to intra-cellular proteins and accelerate folding, assembly and transport of these proteins. It also has molecular chaperones action. The techniques of homology cloning and anchored PCR were used to clong the cyclophilin A (CYPA) gene from Penaeus monodon. We obtained the CYPA'S promoter and genomic DNA sequence from the muscular tissue of Penaeus monodon on basis of designed primers according to the cloned full-length cDNA using genomic walking technology. The results showed after the sequence was analysed, the cDNA sequence contained 495 bp ORF which could translate a putative peptide of 164 amino acids,31 bp 5 UTR and 308 bp 3. UTR. The CYPA genomic DNA amplified from Penaeus monodon genomic library was 3181bp which consists of 1173 bp promoter region, one 426 bp intron and two exons sequences (101 bp, 399 bp). The CYPA genomic DNA has apparente promoter region in the 5 UTR upstream including a GC box and two CAAT box, it lso contains AP1 and CRE regulatory element, all these typical characteristics are accord with eukaryon promoter region, these characteristic conform that our cloned gene belongs to the family of PPlase.