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Proceedings of the 21st Annual Meeting of the European Society for Animal Cell Technology (ESACT), Dublin, Ireland, June 7-10, 2009 pp 47–51Cite as

Analysis of Protein Expression via Alternate 3’ Untranslated Region (UTR) Signals Through the Use of Site Specific Recombination

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Part of the book series: ESACT Proceedings ((ESACT,volume 5))

Abstract

Chinese Hamster Ovary (CHO) cells remain the workhorse of the biopharmaceutical industry. We used site specific recombination in CHO cells to evaluate the post-translational effects of different poly-A signals. Recombinase (Flp) assisted generation of CHO pools show a significant difference in intra-clonal variation within in the pools. Moreover, the use of FRT/Flp resulted in an isogenic population which has allowed accurate correlation between mRNA levels and recombinant protein yields. We identified a human derived 3’UTR which generated a higher level of mRNA when compared to the more commonly used SV40 poly-A, the corresponding recombinant protein levels was found to be independent of the transcript levels.

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References

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Acknowledgements

Robert Simpson from the Real-time PCR facility at the School of Chemical and Molecular Biosciences at the University of Queensland for his assistance with qRT-PCR and Joe Codamo from the AIBN for his help with the poster and qRT-PCR work.

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Authors and Affiliations

  1. Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD, 4072, Australia

    Jeff Jia Cheng Hou, Michael Song & Trent P. Munro

  2. NCRIS Biologics Facility, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, QLD, 4072, Australia

    Peter P. Gray

  3. Acyte Biotech Pty Ltd, Brisbane, QLD, 4072, Australia

    Peter P. Gray

Authors
  1. Jeff Jia Cheng Hou
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  2. Michael Song
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  3. Trent P. Munro
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  4. Peter P. Gray
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Corresponding author

Correspondence to Trent P. Munro .

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Editors and Affiliations

  1. , University College Dublin, National Institute of Bioprocessing Rese, Engineering Building, Belfield, Dublin, Ireland

    Nigel Jenkins

  2. , National Institute for Cellular Biotechn, Dublin City University, Dublin, Ireland

    Niall Barron

  3. Tecnologica (IBET), ITQB, Instituto de Biologia Experimental e, Oeiras, Portugal

    Paula Alves

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© 2012 Springer Science+Business Media B.V.

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Hou, J.J.C., Song, M., Munro, T.P., Gray, P.P. (2012). Analysis of Protein Expression via Alternate 3’ Untranslated Region (UTR) Signals Through the Use of Site Specific Recombination. In: Jenkins, N., Barron, N., Alves, P. (eds) Proceedings of the 21st Annual Meeting of the European Society for Animal Cell Technology (ESACT), Dublin, Ireland, June 7-10, 2009. ESACT Proceedings, vol 5. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-0884-6_8

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