Abstract: To explore the effects of hTRX and PR39 as target genes on cerebral ischemic disease, and to construct a fusion gene of hTRXPR39 cDNA. MethodsThe forward and reverse primers of PR39 were designed and synthesized. By means of PCR, the fragment encoding PR39 was gained, including EcoR721 and BamH I restriction enzyme sites, and the new hTRX cDNA including EcoR721 and EcoRI restriction enzyme sites was also gained. Then the synthesized fragments were easily cloned into vector pGEMT. The positive clone was identified by restriction enzymes, and then the cloned amplified fragments were sequenced by the dideoxymediated chaintermination method. The cloned hTRX and PR39 cDNA were compared with the GeneBank sequence by the DNASIS. After pGEMThTRX and pGEMTPR39 were both digested by BamH I and EcoRI, the PR39/BamHI, EcoRI was cloned into the recombinant vector pGEMThTRX/BamHI,EcoRI .The recombinant vector pGEMThTRXPR39 was gained. ResultsSequences of modified hTRX and PR39 were consistent with those of the gene bank. After analyzing the ORF of the cloned hTRXPR39 cDNA by the DNASIS, we found that amino acids encoded by the cloned hTRXPR39 cDNA were identical to published results. ConclusionThe sequence of hTRX including EcoR721 and EcoRI restriction enzyme sites and sequence of PR39 including EcoR721 and BamH I restriction enzyme sites were successfully obtained by PCR. Also, the recombinant vector pGEMThTRXPR39 was successfully constructed.

Key words: Brain ischemia, Thioredoxin, gene fusion, Antibacterial peptide PR39