小鼠新基因克隆及功能预测

Molecular Cloning and Biochemical Function Prediction of A New Gene from Mouse

该研究克隆了一个可能与小鼠核酸结合相关的新基因,并对其表达模式和预期编码蛋白质功能进行了预测分析。该研究采用启动子捕获技术获得了具有启动子活性的基因组片段,结合生物信息学方法预测得到该启动子活性片段下游基因。采用RT-PCR(reverse transcription PCR)技术扩增得到相应基因,其c DNA为1 504 bp,完整开放阅读框(open reading frame,ORF)为540 bp,编码179个氨基酸。同源性分析表明,该基因与褐鼠(Rattus norvegicus)核酸结合蛋白和黑猩猩(Pan troglodytes)RNA结合蛋白的基因序列分别有78%和40%的相似性。利用实时荧光定量PCR技术分析了不同组织和不同生理、病理状态下该基因的表达水平。结果表明,该基因在带瘤小鼠各组织中的表达水平均高于健康小鼠相应组织,提示该基因可能参与了肿瘤的发生、发展。

In this study, a new gene coding putative nucleic acid binding protein from mouse was cloned. The gene expression pattern and its predicted protein function had also been discussed. By utilizing promoter-trap technology, a DNA fragment with promoter activity was obtained from mouse genomic DNA. A putative gene located in the downstream of this DNA fragment was predicted by bioinformatics analysis. The predicted gene was 1 504 bp in length with an open reading frame of 540 bp. The predicted open reading frame was confirmed by reverse transcription PCR（RT-PCR）. Homology analysis with other species showed that the sequence of this gene shares similarities with the nucleic acid binding protein from Rattus norvegicus and RNA binding protein from Pan troglodytes of 78% and 40%, respectively. The gene expression properties in diverse tissue types under different physiological status or pathological status were studied by quantitative Real-time PCR（q RT-PCR）. As in all the tissue types tested from mice bearing tumor, the expression levels of this gene are much higher than that in healthy mice. These results indicate that this new gene is possibly involved in tumorigenesis.