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An Epitope Tagging Vector for Gene Expression in Mammalian Cells

rotein, composed of FLAG-luciferase-c-myc, is easily detected by Western blot analysis.

In order to demonstrate G418 resistance in the pCMV-Tag1 vector, we used Stratagenes Mammalian Transfection Kit to obtain stable CHO cell lines containing either the pCMV-Tag1 vector or the pCMV-Tag1 vector with the luciferase insert. For the cells transfected with the pCMV-Tag1 vector containing the luciferase gene, the luciferase assay was used to verify the presence of the fusion protein (data not shown).

Conclusions

The pCMV-Tag1 expression vector incorporates the small and highly immunoreactive FLAG and c-myc epitopes into constructs for N-terminal, C-terminal and internal tagging. These tags eliminate the need for raising specific antisera to study a target gene. The epitope tags can be easily detected in transfected cells using well-characterized, commercially available antibodies. The pCMV-Tag1 vector offers a fast, versatile and reliable method for analyzing the function of gene products in vivo.

Acknowledgments

We would like to thank Denise Wyborski, Cathy Chang, Xu Li, Chao-Feng Zheng, Wei-Ping Yang, Phyllis Frosst and the members of the Gerace lab at TSRI for suggestions, discussion and materials.

REFERENCES

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