Aim To obtain the whole gene represented by the expressed sequence tag (EST)AW918640 and analyse its elementary characteristic. Methods Alignment, assembling and other bioinformatics programs were used to perform in silicon cloning form GeneBank data-base. RT-PCR and gene sequencing were performed to identify it. Localization in chromosome, sequence of amino acid and functional domain were identified by map viewer, ExPASy and SMART software respectively. Results AW918640 is an EST sequence that expressed highly at 3 h after myocardial ischemic preconditioning in rat. We got the whole nucleotides sequence represented by AW918640 using in silicon cloning. The result from gene BLAST in GeneBank suggested that the whole nucleotides sequence express a new gene. We named this novel gene myocardial ischemic preconditioning associated gene-5 (MIP5, GenBank accession number: AY553870). Furthermore, the nucleotide sequence of MIP5 was confirmed by RT-PCR and sequencing. MIP5 contained a single open reading frame (ORF)of 909 base pairs with the potential to encode a protein of 302 residues containing 6 kelch repeat motif predicted by map viewer, ExPASy and SMART software respectively. Conclusion MIP5 is a novel gene which is correlative with myocardial ischemic preconditioning in rat.