Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

Expression of exogenous protein in the egg white of transgenic chickens

Nature Biotechnology volume 20, pages 396–399 (2002)Cite this article

Abstract

Using a replication-deficient retroviral vector based on the avian leukosis virus (ALV), we inserted into the chicken genome a transgene encoding a secreted protein, β-lactamase, under the control of the ubiquitous cytomegalovirus (CMV) promoter. Biologically active β-lactamase was secreted into the serum and egg white of four generations of transgenic chickens. The expression levels were similar in successive generations, and expression levels in the magnum of the oviduct were constant over at least 16 months in transgenic hens, indicating that the transgene was stable and not subject to silencing. These results support the potential of the hen as a bioreactor for the production of commercially valuable, biologically active proteins in egg white.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Vector organization and overview of transgenic lineages. (A) Schematic of NLB-CMV-BL.
Figure 2: β-lactamase in the eggs of G2 and G3 hens bred from rooster 4133.
Figure 3: Western blot analysis of β-lactamase in the egg white of transgenic hens.

Similar content being viewed by others

References

  1. Clark, A.J. et al. The molecular manipulation of milk composition. Genome 31, 950–955 (1989).

    Article  CAS  Google Scholar 

  2. Devinoy, E. et al. High level production of human growth hormone in the milk of transgenic mice: the upstream region of the rabbit whey acidic protein (WAP) gene targets transgene expression to the mammary gland. Transgenic Res. 3, 79–89 (1994).

    Article  CAS  Google Scholar 

  3. Ebert, K.M. et al. Transgenic production of a variant of human tissue-type plasminogen activator in goat milk: generation of transgenic goats and analysis of expression. Biotechnology (NY) 9, 835–838 (1991).

    CAS  Google Scholar 

  4. Velander, W.H. et al. High-level expression of a heterologous protein in the milk of transgenic swine using the cDNA encoding human protein C. Proc. Natl. Acad. Sci. USA 89, 12003–12007 (1992).

    Article  CAS  Google Scholar 

  5. Raju, T.S., Briggs, J.B., Borge, S.M. & Jones, A.J. Species-specific variation in glycosylation of IgG: evidence for the species-specific sialylation and branch-specific galactosylation and importance for engineering recombinant glycoprotein therapeutics. Glycobiology 10, 477–486 (2000).

    Article  CAS  Google Scholar 

  6. Love, J., Gribbin, C., Mather, C. & Sang, H. Transgenic birds by DNA microinjection. Biotechnology (NY) 12, 60–63 (1994).

    CAS  Google Scholar 

  7. Carsience, R.S., Clark, M.E., Verrinder Gibbins, A.M. & Etches, R.J. Germline chimeric chickens from dispersed donor blastodermal cells and compromised recipient embryos. Development 117, 669–675 (1993).

    CAS  PubMed  Google Scholar 

  8. Salter, D.W. et al. Gene insertion into the chicken germ line by retroviruses. Poult. Sci. 65, 1445–1458 (1986).

    Article  CAS  Google Scholar 

  9. Bosselman, R.A. et al. Replication-defective vectors of reticuloendotheliosis virus transduce exogenous genes into somatic stem cells of the unincubated chicken embryo. J. Virol. 63, 2680–2689 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Thoraval, P. et al. Germline transmission of exogenous genes in chickens using helper-free ecotropic avian leukosis virus–based vectors. Transgenic Res. 4, 369–377 (1995).

    Article  CAS  Google Scholar 

  11. Harvey, A.J., Speksnijder, G., Baugh, L.R., Morris, J.A. & Ivarie, R. Consistent production of transgenic chickens using replication deficient retroviral vectors and high-throughput screening procedures. Poult. Sci. 81, 202–212 (2002).

    Article  CAS  Google Scholar 

  12. Petropoulos, C.J., Payne, W., Salter, D.W. & Hughes, S.H. Using avian retroviral vectors for gene transfer. J. Virol. 66, 3391–3397 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Moore, J.T., Davis, S.T. & Dev, I.K. The development of β-lactamase as a highly versatile genetic reporter for eukaryotic cells. Anal. Biochem. 247, 203–209 (1997).

    Article  CAS  Google Scholar 

  14. Cosset, F.L. et al. Improvement of avian leukosis virus (ALV)–based retrovirus vectors by using different cis-acting sequences from ALVs. J. Virol. 65, 3388–3394 (1991).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Cosset, F.L. et al. A new avian leukosis virus–based packaging cell line that uses two separate transcomplementing helper genomes. J. Virol. 64, 1070–1078 (1990).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Cosset, F.L. et al. Packaging cells for avian leukosis virus–based vectors with various host ranges. J. Virol. 66, 5671–5676 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Fadly, A.M. & Smith, E.J. Isolation and some characteristics of a subgroup J–like avian leukosis virus associated with myeloid leukosis in meat-type chickens in the United States. Avian Dis. 43, 391–400 (1999).

    Article  CAS  Google Scholar 

  18. Schaefer-Klein, J. et al. The EV-O-derived cell line DF-1 supports the efficient replication of avian leukosis-sarcoma viruses and vectors. Virology 248, 305–311 (1998).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Robert Harvey for the β-lactamase plasmid and Henry Marks and the University of Georgia Poultry Science Department for access to poultry facilities. We thank Wei Hu, Lyn Olliff, Michael Morgan, Jeff Rapp, Sara Mathews, Heath Sellers, and Tom Beacorn for technical assistance. This work was supported in part by research grants to R.I. from the Georgia Research Alliance, the Applied Technology Development Corp., and AviGenics, Inc.

Author information

Author notes

  1. Gordon Speksnijder

    Present address: AviGenics, Inc., Georgia BioBusiness Center, 111 Riverbend Rd., Athens, GA, 30605

  2. Larry R. Baugh

    Present address: Department of Molecular and Cellular Biology, Harvard University, 16 Divinity Street, Cambridge, MA, 02138

Authors and Affiliations

  1. AviGenics, Inc., Georgia BioBusiness Center, 111 Riverbend Rd., Athens, 30605, GA

    Alex J. Harvey

  2. Department of Genetics, University of Georgia, Athens, 30602-7223, GA

    Gordon Speksnijder, Larry R. Baugh, Julie A. Morris & Robert Ivarie

Authors
  1. Alex J. Harvey
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gordon Speksnijder
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Larry R. Baugh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Julie A. Morris
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert Ivarie
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alex J. Harvey.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Harvey, A., Speksnijder, G., Baugh, L. et al. Expression of exogenous protein in the egg white of transgenic chickens. Nat Biotechnol 20, 396–399 (2002). https://doi.org/10.1038/nbt0402-396

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nbt0402-396

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing