人UBF新基因功能的初步研究

发布时间：2018-05-05 19:36

本文选题：UBF基因 + 生物信息学　；参考：《中国人民解放军军事医学科学院》2005年硕士论文

【摘要】：UBF (GenBank注册名:UBF fl-AF294842)基因来源于本室人胎肝cDNA文库大规模测序的EST库(HA2180),经进一步测序获得。根据该基因核苷酸序列的5’-端有高比例GC特征,在起始密码子前的同相位上有连续两个终止密码子,3’-端有poly-A尾的特点,认为它是一条全长cDNA序列。基因组结构分析证实UBF由一个外显子组成,不含任何内含子。电子PCR染色体定位提示,UBF定位于人染色体19q13.4。核苷酸和氨基酸序列同源性比较结果显示,UBF基因与泛素结合酶家系成员高度同源。开放阅读框(ORF Finder)软件分析结果提示,该基因可能含有两个开放阅读框(ORF),分别编码89个和105个氨基酸,两ORF之间相隔30个核苷酸,包括前一个ORF的终止密码子。人胎肝和HL-60细胞mRNA的RT-PCR产物直接测序结果证实了该基因的天然属性及中间终止密码子的客观实在性;基因转染及体外翻译实验结果提示UBF基因中第一个ORF是编码序列,其后包括第二个ORF在内的序列为3’端非翻译区。对UBF基因功能的进一步研究发现,将该基因导入到COS7细胞内进行高表达时,与对照组相比,可导致COS7细胞的凋亡;为了进一步探讨UBF的功能及作用机理,后期实验中引入Tet-on/off基因表达系统,研究结果显示,UBF在CHO细胞内稳定表达后,出现几乎百分之百的多倍体细胞。上述实验结果给UBF基因的功能研究提供了一定线索,具体作用机理尚待进一步研究。UBF是一条来源于高度进化的人类的胎肝组织的新基因,注册至今尚无相关文献报道,实验中遗留的许多问题还有待于进一步的分析和证明,从而揭开UBF基因的神秘面纱。
[Abstract]:The UBF GenBank registered name: UBF fl-AF294842) was obtained from EST library HA2180, which was sequenced on a large scale in human fetal liver cDNA library. According to the high proportion GC characteristics of the nucleotide terminal of the nucleotide sequence and the poly-A tail in the same phase of the starting codon, it is considered that it is a full-length cDNA sequence. Genomic structural analysis confirmed that UBF consists of an exon and does not contain any introns. The electron PCR chromosome localization indicates that UBF is located on human chromosome 19q13.4. The homology of nucleotide and amino acid sequences showed that the UBF gene was highly homologous to the family members of ubiquitin binding enzyme. The results of ORF Finder software analysis suggested that the gene might contain two ORFUs encoding 89 amino acids and 105 amino acids, respectively, with 30 nucleotides separated between the two ORF, including the termination codon of the previous ORF. The results of direct sequencing of mRNA RT-PCR products from human fetal liver and HL-60 cells confirmed the nature of the gene and the objective reality of the intermediate termination codon. The results of gene transfection and in vitro translation showed that the first ORF of the UBF gene was a coding sequence. The sequence, including the second ORF, was a 3'- end untranslated region. Further studies on the function of UBF gene showed that when the gene was introduced into COS7 cells for high expression, it could induce apoptosis of COS7 cells compared with the control group, in order to further explore the function and mechanism of UBF. Tet-on/off gene expression system was introduced in the later experiment. The results showed that almost 100% of polyploid cells appeared after stable expression in CHO cells. These results provide some clues for the functional study of UBF gene. The specific mechanism of the study remains to be further studied. UBF is a new gene derived from highly evolved human fetal liver tissue. Many of the problems left over in the experiment need to be further analyzed and proved to reveal the mystery of the UBF gene.
【学位授予单位】：中国人民解放军军事医学科学院
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R346

【参考文献】