大鼠心肌缺血预适应相关基因5的克隆与特性分析

目的采用生物信息学分析表达序列标签AW918640以获得其所代表基因的全长序列,并对该基因进行初步的特性分析。方法利用基因银行(GeneBank)数据库,通过BLAST比对等生物信息学方法进行电子克隆,采用逆转录聚合酶链反应和基因测序等技术进行鉴定,采用mapviewer软件进行染色体定位,ExPASy和SMART软件对获得的全长基因进行翻译和蛋白质功能结构分析。结果AW918640(基因银行登录号)为一个在大鼠心肌缺血预适应后3h表达升高的表达序列标签序列,通过电子克隆得到其所代表基因的全长,且为一新基因,经逆转录取合酶链反应和基因测序证实后,将其命名为心肌缺血预适应相关基因5(MIP5),并在基因银行数据库上登录,登录号为AY553870。心肌缺血预适应相关基因5最大开放读码框为909bp,编码302个氨基酸,含有6个kelch重复基序。结论心肌缺血预适应相关基因5为一与大鼠心肌缺血预适应相关的新基因。

Cloning and Characterization of Myocardial Ischemic Preconditioning Associated Gene five (MIP5) in the Rat

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.