一条大鼠睾丸新基因的生物信息学分析及真核表达

Bioinformatic Analysis and Eukaryotic Expression of a New Rat Testicular cDNA Gene

目的:探讨大鼠睾丸组织一条新基因的生物信息学特征和真核表达。方法:构建pEGFP-N1载体的融合质粒进行真核表达,利用生物信息学手段分析基因和蛋白功能。结果:生物信息学分析表明RSA14-44的编码区序列与人类及鼠源RAS同源基因家族核酸序列达到85%以上的同源性;RSA14-44蛋白没有典型的跨膜结构域,也没有典型的N末端信号肽;与人类RhoA蛋白序列达到了89%的同源且具有Rho家族成员的GAAX盒和p-loop结构的基序特征;RSA14-44蛋白大部分氨基酸序列与Rho家族7个已知结构域高度同源;RSA14-44基因真核表达定位于细胞质。结论:RSA14-44基因真核表达定位于CHO-K1细胞质,;编码蛋白质与Rho家族同源性高,为进一步研究其生物学功能提供参考。

Objective：To investigate the bioinformatic analysis and eukaryotic expression of a new rat testicular cDNA gene.Methods：The gene was amplified by PCR from genomic DNA of rat testis Genes and was eukaryotic expressed by a recombinant expression plasmid pEGFP-N1-14,the structure and the function of cloning genes were predicted by bioinformatics.Results：The homologue of the RSA14-44 gene between the rat and human chromosome RAS family DNA sequence was 85% with and the homologue of this gene coding protein was 89% with human family RhoA.There were typically GAAX box and p-loop sequence at the coding region;There were no typical membrane spaning domain and terminal signal peptide.However,the RSA14-44 amino acid sequence had high homologous with the seven known structural domain of family Rho.The localization for RSA14-44 in CHO cells was observed in cytoplasm.Conclusion：The coding protein of rat gene was localized in cytoplasm and it was high homologous with Rho family.