Mouse IL-33 gene cDNA cloning and eukaryotic expression vector and.doc

 PAGE \* MERGEFORMAT 10 Mouse IL-33 gene cDNA cloning and eukaryotic expression vector and Of: Zhang, Cheng Xiaogang, Xu Lian Hui, Li Xin, ON TRADITIONAL, Zhao Bo Cheng, Ren Fei, Li Han-gun, Liu Ping [Abstract] Objective To clone the mouse IL-33 gene (mIL-33) cDNA eukaryotic expression vector, and transfected EL-4 cells to detect expression. Extracted from mouse brain and lung total RNA, by reverse transcription - polymerase chain reaction (RT-PCR amplification of mouse IL-33 gene cDNA, inserted into pcDNA-3.1 was digested eukaryotic expression vector pcDNA-mIL-33, recombinant plasmid transfected EL-4 cells, RT-PCR and ELISA assay gene expression. Results pcDNA-mIL-33 in the inserted DNA sequencing results with the mIL-33 cDNA sequences of the same plasmids were transfected into EL-4 cells were detected in the corresponding gene expression. Conclusion cloned mouse IL- 33 gene cDNA, and construct its eukaryotic expression plasmid. [Keywords:] IL-33, gene cloning, eukaryotic expression Abstract: Objective To clone and construct the eukaryotic expression plasmids containing mouse IL-33 cDNA and investigate their expression. Methods Total RNA was extracted from mouse lung and brain tissues to amplify mouse interleukin-33 (mIL-33) cDNA by reverse transcription-polymerase chain reaction (RT-PCR). The eukaryotic expression plasmids pcDNA-mIL-33 was constructed with the insertion of enzyme-digested pcDNA-3.1 (+) vector to construct eukaryotic expression vector pcDNA-mIL-33. The recombinant plasmids were transfected into EL-4 cells and the expression of target gene was detected by RT-PCR and ELISA. Results The inserted DNA sequence in pcDNA-mIL-33 was identical to the mIL-33 cDNA, which was verified correctly by sequencing. The corresponding gene expression was detected in the recombinant plasmid-transfected EL-4 cells.Conclusion The mouse IL-33 cDNA was successfully cloned and its eukaryotic expression plasmids were constructed. Keywords:: IL-33; gene