高凝状态大鼠主动脉表达上调新基因HCR2的克隆及生物信息学分析

目的 获得高凝状态大鼠主动脉上调表达新基因HCR2的全长cDNA，并对该基因进行生物信息学分析。方法 从高凝大鼠主动脉中表达显著上调的序列标签（EST，GenBank登录号为BQ901227）出发，利用cDNA末端快速扩增（rapid amplification of cDNA end，RACE）和巢式PCR技术扩增其全长cDNA，利用NCBI的数据库对其进行生物信息学分析。结果 成功地获得了HCR2的全长cDNA，在GenBank中的登录号为AY234417。用RT-PCR证实HCR2是高凝大鼠表达上调基因。生物信息学分析显示，该基因cDNA序列全长为1275bp，定位于大鼠染色体4q11，编码由78个氨基酸组成、相对分子质量为8841．7、pI为8．59的蛋白质；无信号肽序列和跨膜序列，很可能是一种核蛋白；与已知蛋白无明显同源性。结论 HCR2的成功克隆为进一步研究其生物学功能及其在高血凝的发生、发展中的作用奠定了坚实基础。

Cloning and Bioinformatic Analysis of a Novel Gene HCR2 Up-regulated in Aorta of Hypercoagulable Rat

Objective The aim of this study was to clone the full-length cDNA of HCR2 up-regulated in aorta of hypercoagulable rat and make the relevant bioinformatic analysis. Methods Rapid amplification of cDNA end (RACE) and nested PCR technique was used to amplify the full-length cDNA of HCR2 from an EST(GenBank accession number BQ901227)which was significantly up-regulated in hypercoagulable rat. Results The full-length cDNA of HCR2 has been obtained (GenBank accession number AY234417). Semi-quantitative RT-PCR demonstrated that HCR2 is up-regulated in aorta of hypercoagulable rat. Bioinformatic analysis showed that HCR2 gene is located in rat chromosome 4q11. The full-length cDNA of HCR2 is 1275 bp, coding for a 78 amino acids polypeptide with a theoretical molecular weight of 8841.7 and isoelectric point of 8.59. This protein has no signal peptide and transmembrane sequence. It may be a nuclear protein. It shares no significant homology with any known protein. Conclusion The successful cloning of HCR2 has laid a solid foundation for further study of its function and its possible role in the occurrence and development of hypercoagulability.