UBC家族新成员UBF基因的组织表达谱研究

目的和方法:提取人胎肝和HL-60细胞总RNA,采用RT-PCR的方法,从中扩增UBF基因,并对扩增产物进行测序,从而证实UBF基因的天然存在性;采用原位杂交的方法研究UBF在5月龄胎儿的不同组织及HL-60细胞中的表达谱及亚细胞定位.结果:从胎肝和HL-60细胞中均扩增得到了完整的UBF基因,且其序列与注册序列完全一致;原位杂交结果显示UBF在人胚胎多种组织中表达,其中胎骨骼肌表达最强,胎肾最弱.结论:UBF基因,作为Ubc家族的一个新成员,在人胚胎的骨骼肌、肝脏、肾脏、心脏和肺中均有表达,且骨骼肌中的表达最强.

Expression profiling of UBF, a novel member of UBC family

From large-scale sequence of human fetal liver cDNA library,we have obtained a full-length cDNA from an EST after further sequen- cing . It has been demonstrated by the alignment comparison with data base available that it is a novel member of Ubc family and got the number from GeneBank: UBF-Fl AF 294842. Aim and Methods: To demonstrate its authenticity,UBF was amplified from the total RNA of human fetal liver and HL-60 cell line using RT-PCR,and the PCR products were further sequenced and compared with the original UBF sequence. To evaluate the expression level and subcellular location of UBF in human multiple tissues,in situ hybridization was carried out on the frozen section of human fetal multiple tissues and HL-60 cell line with DIG-labeled UBF cDNA probes. Results: The experimental results of RT-PCR and sequencing showed that the sequence of RT-PCR products were the same as the original UBF. The experimental results of in situ hybridization showed that UBF was expressed widely by human multiple fetal tissues and the expression level were very high in HL-60 cells. Conclusion: It is suggested that the special structure of UBF is authentic,and the expression profiling research of UBF shows that UBF is expressed widely by human multiple fetal tissues and the expression level is very high in HL-60 cells,implying that UBF plays the important function in the developing tissues and leukemia cells. It is also suggested that UBF may be functionally related with the nucleic-involving cellular activities based on the results of sub-cellular localizations.