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Published on: October 30, 2018
[Construction of a fusion gene expression vector containing mouse Ipr1 and EGFP and its expression in murine
Na Li1, Dao-yin Zhu, Ru-xiu Tie
1Department of Immunology, Chongqing University of Medical Sciences, Chongqing 400016, China. friend_lina@163.com
Aim:
To construct an eukaryotic expression vector containing the fusion gene of mouse Ipr1 and EGFP and to study its expression in murine macrophage RAW264.7.
Methods:
The coding sequence of Ipr1 gene was amplified from the total RNA of C57BL/6J mouse thymus by RT-PCR. The gene was cloned into pEGFP-C1 and the recombinant plasmid was identified by PCR, restrict endonuclease digestion and sequencing. Then the pEGFP-Ipr1 was transiently transfected into RAW264.7. The expression of Ipr1 gene and fusion protein was detected by RT-PCR and laser scanning confocal microscopy.
Results:
The whole coding sequence of Ipr1 was successfully amplified. The recombinant plasmid was identified by PCR, restrict endonuclease digestion and sequencing. The fusion protein was successfully expressed in the targeted cells and its localization was in nucleus.
Conclusion:
The eukaryotic expression vector pEGFP-Ipr1 has been successfully constructed. The fusion protein can be expressed in murine macrophage RAW264.7 and located in nucleus.
