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The contribution of the C terminal SPRY domain of TRIM22 to its own transcription, translation, and subcellular localization |
GAO Bo; DUAN Zhi-Jian; XU Wei; XIONG Si-Dong |
Department of Immunology, Shanghai Medical College, Institute for Immunobiology, Fudan University, Shanghai 200032, China |
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Abstract In oeder to investigate the effects of C terminal SPRY domain of TRIM22 on its transcription, translation and subcellular localization, SPRY domain was deleted (ΔSPRY) and amplified through polymerase chain reaction (PCR) and was then inserted into the eukaryotic expression plasmid, pcDNA3.1. Plasmids expressing wild type TRIM22 or TRIM22-ΔSPRY were transfected into HepG2 cells. The mRNA expression level was determined by semiquantitative reverse transcriptase PCR (RT-PCR), the protein expression level was determined by Western blot analysis, and the subcellular localization was determined by immunofluorescence staining. The results showed that plasmid expressing TRIM22-ΔSPRY was successfully constructed. After transfection into HepG2 cells, the TRIM22-ΔSPRY showed no difference from the wild type TRIM22 at both mRNA and protein levels. However TRIM22-ΔSPRY was localized exclusively to the cytoplasm of HepG2 cells in contrast to the nuclear localization of wild type TRIM22. These results may be useful for studying the role of SPRY domain in TRIM22-mediated antiviral activities.
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Received: 04 August 2009
Published: 25 March 2010
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Corresponding Authors:
XIONG Si-Dong
E-mail: sdxiong@shmu.edu.cn
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