title: [Identification and functional analysis of a testis-specific E3 ubiquitin ligase gene Rnf148 in mouse].
abstract: OBJECTIVE
  To investigate the temporal and spatial features of mouse Rnf148 gene expression and the function of RING finger domain of Rnf148 protein.
  
  
  METHODS
  The whole RNA was extracted from different tissues of adult mice, embryo in four developmental stages, and testes of postnatal mice respectively. RT-PCR and Northern blotting analysis were used to investigate the expression of Rnf148 gene in the above tissues. The in vitro expression vector for GST-Rnf148 fused protein was constructed, which encompassing the entire RING domain of Rnf148 protein. GST-Rnf148 fused protein was expressed in Escherichia coli. BL21(DE3) cells and purified with glutathione-sepharose 4B. In vitro ubiquitination assay was performed to analyze whether GST-Rnf148 fused protein possess the function of E3 ubiquitin ligase.
  
  
  RESULTS
  The Mice Rnf148 mRNA expression was only observed in testis, and Northern blotting confirmed that there was only one 1.2 kb mRNA band present in mice testis. Rnf148 mRNA started to appear in the testis of day 21 mice, and then increased dramatically and reached to the highest level in day 25, and continued to express thereafter. GST-Rnf148 fused protein was induced and purified, in vitro ubiquitination reaction showed that the recombinant protein has E3 ubiquitin ligase activity.
  
  
  CONCLUSIONS
  Rnf148 gene is specifically expressed in mice testis.
authors: Department of Medical Genetics & Division of Human Morbid Genomics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors: Yun-Qiang
authors: YQ
authors: Liu
authors: Department of Medical Genetics & Division of Human Morbid Genomics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors: Da-Chang
authors: DC
authors: Tao
authors: Institute of Organ Transplantation, Sichuan Academy of Medical Science, Sichuan Provincial People's Hospital, Chengdu 610061, China.
authors: Shun-Yao
authors: SY
authors: Liao
authors: Department of Medical Genetics & Division of Human Morbid Genomics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors: Yuan
authors: Y
authors: Yang
authors: Department of Medical Genetics & Division of Human Morbid Genomics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors: Yong-Xin
authors: YX
authors: Ma
authors: Department of Medical Genetics & Division of Human Morbid Genomics, State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors: Si-Zhong
authors: SZ
authors: Zhang
mesh_terms: D000818:Animals; D004926:Escherichia coli; D015870:Gene Expression; D008297:Male; D051379:Mice; D012333:RNA, Messenger; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013737:Testis / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D011994:Recombinant Proteins; D044767:Ubiquitin-Protein Ligases