Aim To clone the upregulated expressed gene during rat myocardium ischemic preconditioning. Methods New gene was in silicon cloned using exon finding, blast alignment, assembling program and so on. Reverse transcriptase polymerase chain reaction (RT-PCR) assay confirmed the open reading frame (ORF) of new gene. Results Rat heterogeneous nuclear ribonucleoproteins (hnRNP) A2/B1 gene was cloned by bioinformatics methods. The product of RT-PCR was 1.1 kb, which was identical to the predicted length. The ORF of rat hnRNPA2/Bl gene was 1 026 bp, which was composed of ten exons and encoded 341 amino acids. The coding sequence showed 92% and 95% identity respectively in 1 026 bp overlap with that of human and mouse, and the predicted polypeptide showed almost 100% identity with that of human and mouse. Conclusions The results showed that hnRNPA2/Bl gene was highly conserved among various species. It maybe plays an important role in endogenous protection of ischemic preconditioning.