Abstract
Achieving high expression levels of recombinant human serum albumin (HSA) for purification is a solution for the large amount of plasma-derived HSA needed in therapeutic applications. Here, we employed phiC31 integrase system and chicken hypersensitive site-4 (cHS4) insulators to construct a HSA expression vector for high-level HSA expression. The phiC31 integrase system mediated efficient transgene integration in bovine mammary epithelial cells (bMECs). A preferred pseudo attP site, which had 38 % identity with the 39 bp wild-type attP sequence, was detected in six out of 55 bMEC colonies. Addition of the cHS4 insulator to the phiC31 integrase system resulted in 8–20-fold increases of HSA expression compared with that of using integrase alone. Moreover, the reverse-oriented cHS4 insulator in the phiC31 integrase system provided the optimal level of HSA expression in bMECs.
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Acknowledgments
We thank Prof. M. P. Calos and Dr. Yuan Yu for plasmids. We are grateful to Prof. Zekun Guo and Dr. Yongyan Wu for their technical advice. Our work was supported by the National Major Project for Production of Transgenic Breeding (No. 2011ZX08007-004) and the National High Technology Research and Development Program of China (863 Program) (No. 2011AA100303).
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Yan Luo and Jun Liu contributed equally to this work.
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Luo, Y., Liu, J., Liu, Q. et al. Chicken hypersensitive site-4 insulator increases human serum albumin expression in bovine mammary epithelial cells modified with phiC31 integrase. Biotechnol Lett 35, 529–537 (2013). https://doi.org/10.1007/s10529-012-1125-y
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DOI: https://doi.org/10.1007/s10529-012-1125-y