本文選題：UBF基因 + 生物信息學(xué)　； 參考：《中國人民解放軍軍事醫(yī)學(xué)科學(xué)院》2005年碩士論文

【摘要】：UBF (GenBank注冊名:UBF fl-AF294842)基因來源于本室人胎肝cDNA文庫大規(guī)模測序的EST庫(HA2180),經(jīng)進(jìn)一步測序獲得。根據(jù)該基因核苷酸序列的5’-端有高比例GC特征,在起始密碼子前的同相位上有連續(xù)兩個終止密碼子,3’-端有poly-A尾的特點(diǎn),認(rèn)為它是一條全長cDNA序列�；蚪M結(jié)構(gòu)分析證實(shí)UBF由一個外顯子組成,不含任何內(nèi)含子。電子PCR染色體定位提示,UBF定位于人染色體19q13.4。核苷酸和氨基酸序列同源性比較結(jié)果顯示,UBF基因與泛素結(jié)合酶家系成員高度同源。開放閱讀框(ORF Finder)軟件分析結(jié)果提示,該基因可能含有兩個開放閱讀框(ORF),分別編碼89個和105個氨基酸,兩ORF之間相隔30個核苷酸,包括前一個ORF的終止密碼子。人胎肝和HL-60細(xì)胞mRNA的RT-PCR產(chǎn)物直接測序結(jié)果證實(shí)了該基因的天然屬性及中間終止密碼子的客觀實(shí)在性;基因轉(zhuǎn)染及體外翻譯實(shí)驗(yàn)結(jié)果提示UBF基因中第一個ORF是編碼序列,其后包括第二個ORF在內(nèi)的序列為3’端非翻譯區(qū)。對UBF基因功能的進(jìn)一步研究發(fā)現(xiàn),將該基因?qū)氲紺OS7細(xì)胞內(nèi)進(jìn)行高表達(dá)時,與對照組相比,可導(dǎo)致COS7細(xì)胞的凋亡;為了進(jìn)一步探討UBF的功能及作用機(jī)理,后期實(shí)驗(yàn)中引入Tet-on/off基因表達(dá)系統(tǒng),研究結(jié)果顯示,UBF在CHO細(xì)胞內(nèi)穩(wěn)定表達(dá)后,出現(xiàn)幾乎百分之百的多倍體細(xì)胞。上述實(shí)驗(yàn)結(jié)果給UBF基因的功能研究提供了一定線索,具體作用機(jī)理尚待進(jìn)一步研究。UBF是一條來源于高度進(jìn)化的人類的胎肝組織的新基因,注冊至今尚無相關(guān)文獻(xiàn)報(bào)道,實(shí)驗(yàn)中遺留的許多問題還有待于進(jìn)一步的分析和證明,從而揭開UBF基因的神秘面紗。
[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.
【學(xué)位授予單位】：中國人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R346

【參考文獻(xiàn)】

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1 ;The IAP family: endogenous caspase inhibitors with multiple biological activities[J];Cell Research;2000年03期

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本文編號：1849018