Skip to main content
SpringerLink
Log in

Production of human surfactant protein C in milk of transgenic mice

  • Papers
  • Published:
Transgenic Research Aims and scope Submit manuscript

Abstract

Respiratory distress syndrome (RDS), caused by lack of pulmonary surfactant, affects 65 000 infants annually in the USA. Surfactant replacement therapy reduces the morbidity and mortality associated with RDS. Human surfactant protein C (SP-C) is an important component of pulmonary surfactant. To produce human SP-C, a construct using the rat whey acidic protein (WAP) promoter and 3′ untranslated regions to target expression of the human SP-C gene to the mammary gland of transgenic mice was created. WAP/SP-C mRNA expression was detected in all transgenic lines analysed. SP-C was expressed in a copy-number-dependent and integration-site-independent fashion, with levels of expression ranging from 0.01% to 36.0% of the endogenous mouse WAP mRNA, and WAP/SP-C mRNA expresison levels were greater than those of the endogenous mouse lung SP-C mRNA. Expression at the RNA level was specific to the mammary gland and paralleled the endogenous WAP expression pattern during mammary gland development. Expression and secretion of the SP-C protein in the lactating mammary gland was demonstrated by western blots performed on whole milk using an anti-SP-C polyclonal antibody. Immunoreactive proteins of MW 22 and 12–14 kDa appeared only in transgenic milk. The 22 kDa protein represents the proprotein, and the 12–14 kDa is a processed form of SP-C.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Archibald, A., McClenaghan, M., Hornsey, V., Simons, J.P., and Clark, A.J. (1990) High-level expression of biologically active human α1-antitrypsin in the milk of transgenic mice.Proc. Natl Acad. Sci. USA 87, 5178–82.

    PubMed  Google Scholar 

  • Bayna, E.M. and Rosen, J.M. (1990) Tissue-specific, high level expression of the rat whey acidic protein gene in transgenic mice.Nucl. Acids Res. 18, 2977–2985.

    PubMed  Google Scholar 

  • Bühler, T., Bruyere, T., Went, D.F., Stranzinger, G. and Bürki, K. (1989) Rabbit β-casein promoter directs secretion of human interleukin-2 into the milk of transgenic rabbits.Bio/Technology 8, 140–3.

    Article  Google Scholar 

  • Burdon, T., Sankaran, L., Wall, R.J., Spencer, M. and Hennighausen, L. (1991a) Expression of a whey acidic protein transgene during mammary development.J. Biol. Chem. 266, 6909–14.

    PubMed  Google Scholar 

  • Burdon, T., Wall, R.J., Shamay, A., Smith, G.H. and Hennighausen, L. (1992b) Over-expression of an endogenous milk protein gene in transgenic mice is associated with impaired mammary alveolar development and amilchlos phenotype.Mech. Devel. 36, 67–74.

    Article  Google Scholar 

  • Carver, A.S., Dalrymple, M.A., Wright, G., Cottom, D.S., Reeves, D.B., Gibson, Y.H., Keenan, J.L., Barrass, J.D., Scott, A.R., Colman A. and Garner, I. (1993) Transgenic livestock as bioreactors: stable expression of human alpha-1-antitrypsin by a flock of sheep.Bio/Technology 11, 1263–70.

    PubMed  Google Scholar 

  • Clark, A.J. (1992) Prospects for the genetic engineering of milk.J. Cell. Biochem. 49, 121–7.

    Article  PubMed  Google Scholar 

  • Curstedt, T., Johansson, J., Persson, P., Eklund, A., Robertson, B., Lowenadler, B. and Jornvall, H. (1990) Hydrophobic surfactant-associated polypeptides: SP-C is a lipopeptide with two pamitoylated cysteine residues, whereas SP-B lacks covalently linked fatty acyl groups.Proc. Natl Acad. Sci. USA 87, 2985–9.

    PubMed  Google Scholar 

  • Dale, T.C., Krnacik, M.J., Schmidhauser, C., Yang, C.L.Q., Bissell, M.J. and Rosen, J.M. (1992) High-level expression of the rat whey acidic protein gene is mediated by elements in the promoter and 3′ untranslated region.Mol. Cell. Biol. 12, 905–14.

    PubMed  Google Scholar 

  • Ebert, K.M., Selgrath, J.P., DiTullio, P., Denman, P., Memmon, M.A., Schindler, J.E., Monastersky, G.M., Vitale, J. and Gordon, K. (1991) Transgenic production of a variant of human tissue-type plasminogen activator in goat milk. Generation of transgenic goats and analysis of expression.Bio/Technology 9, 835–8.

    Article  PubMed  Google Scholar 

  • Glasser, S.W., Korfhagen, T.R., Perme, C.M., Pilot-Matias, T.J., Kister, S.E. and Whitsett, J.A. (1988) Two SP-C genes encoding human pulmonary surfactant proteolipid.J. Biol. Chem. 263, 10326–31.

    PubMed  Google Scholar 

  • Gordon, K., Lee, E., Vitale, J.A., Smith, A.E., Westphal, H. and Henninghausen, L. (1987) Production of human tissue plasminogen activator in transgenic mice.Bio/Technolgy 5, 1183–7.

    Article  Google Scholar 

  • Greenberg, N.M., Anderson, J.W., Hsueh, A.J.W., Nishimori, K., Reeves, J.J., deAvila, D.M., Ward, D.N. and Rosen, J.M. (1991) Expression of biologically active heterodimeric bovine follicle-stimulating hormone in milk of transgenic mice.Proc. Natl Acad. Sci. USA 88, 8327–31.

    PubMed  Google Scholar 

  • Haagsman, H.P. and Golde, L.M.G. van (1991) Synthesis and assembly of lung surfactant.Annu. Rev. Physiol. 53, 441–64.

    Article  PubMed  Google Scholar 

  • Harlow, E. and Lane, D. (1988)Antibodies: a Laboratory Manual. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Hawgood, S. (1989) Pulmonary surfactant apoproteins: a review of protein and genomic structure.Am. J. Physiol. (Lung Cell Mol. Physiol.) 257, L13–22.

    Google Scholar 

  • Hennighausen, L. (1990) The mammary gland as a bioreactor: production of foreign proteins in milk.Prot. Expr. Purif. 1, 3–8.

    Article  Google Scholar 

  • Hoekstra, R.E., Jackson, J.C., Myers, T.F., Frantz, I.D. III, Stern, M.E., Powers, W.F., Maurer, M., Raye, J.R., Carrier, S.T. and Gunkel, J.H. (1991) Improved neonatal survival following multiple doses of bovine surfactant in very premature neonates at risk of respiratory distress syndrome.Pediatrics 88, 10–8.

    PubMed  Google Scholar 

  • Jobe, A.H. (1993) Pilmonary surfactant therapy.New Eng. J. Med. 328 861–8.

    Article  PubMed  Google Scholar 

  • Lee, K.-F., DeMayo, F.J., Atiee, S.H. and Rosen, J.M. (1988) Tissue-specific expression of the rat β-casein gene in transgenic mice.Nucl. Acids Res. 16 1027–41.

    PubMed  Google Scholar 

  • Lee, K.F., Atiee, S.H. and Rosen, J.M. (1989) Differential regulation of rat β-casein-chloramphenicol acetyltransferase fusion gene expression in transgenic mice.Mol. Cell. Biol. 9, 560–5.

    PubMed  Google Scholar 

  • Lewis, J.F. and Jobe, A.H. (1993) Surfactant and the adult respiratory distress syndrome.Am. Rev. Respir. Dis. 147, 218–33.

    PubMed  Google Scholar 

  • Li, B., Greenberg, N., Stephens, L.C., May, R., Medina, D. and Rosen, J.M. (1994) Preferential overexpression of a 172Arg-Leu mutant p53 in the mammary gland of transgenic mice results in altered lobuloalveolar development.Cell Growth Diff. 5, 711–21.

    PubMed  Google Scholar 

  • Leichty, E.A., Donovan, E., Purohit, D., Gilhooly, J., Feldman, B., Noguchi, A., Denson, S.E., Sehgal, S.S., Gross, I. and Stevens, D. (1991) Reduction of neonatal mortality after multiple dose of bovine surfactant in low birthweight neonates with respiratory distress syndrome.Pediatrics 88, 19–28.

    PubMed  Google Scholar 

  • McKnight, R.A., Shamay, A., Sankaran, L., Wall, R. and Hennighausen, L. (1992) Matrix-attachment regions can impart position-independent regulation of a tissue-specific gene in transgenic mice.Proc. Natl Acad. Sci. USA 89 6943–7.

    PubMed  Google Scholar 

  • Meade, J.C., Gates, L., Kacy, E. and Lonberg, N. (1990) Bovine αs1-casein gene sequences direct high level expression of active human urokinase in mouse milk.Bio/Technology 8, 443–6.

    Article  PubMed  Google Scholar 

  • Pittius, C.W., Hennighausen, L., Lee, E., Westphal, H., Nichols, E., Vitale, J. and Gordon, K. (1988) A milk protein gene promoter directs the expression of human tissue plasminogen activator cDNA to the mammary gland in transgenic mice.Proc. Natl Acad. Sci. USA 85, 5874–8.

    PubMed  Google Scholar 

  • Reynolds, M.S. and Wallander, K.A. (1989) Use of surfactant in the prevention and treatment of neonatal respiratory distress syndrome.Clin. Pharm. 8 559–76.

    PubMed  Google Scholar 

  • Sall, J., Ng, K., Hecht, M. and Lehnan, A. (1992) JMP software for statistical visualization of the Apple Macintosh. In Cary, NC: SAS Institute Inc.

    Google Scholar 

  • Saugstad, O.D., Halliday, H.L., Robertson, B. and Speer, C.P. (1993) Replacement therapy with porcine natural surfactant-current status and future challenges.Biol. Neonate 64, 269–78.

    PubMed  Google Scholar 

  • Schagger, H. and Jagow, G. von (1987) Tricine sodium-dodecyl sulfate-polyacrylamide gel electrophoresis.Anal. Biochem. 166, 368–79.

    PubMed  Google Scholar 

  • Schwartz, R.M., Luby, A.M., Scanlon, J.W. and Kellogg, R.J. (1994) Effect of surfactant on morbidity, mortality, and resource use in newborn infants weighing 500 to 1500 g.New Eng. J. Med. 330, 1476–80.

    Article  PubMed  Google Scholar 

  • Stults, J.T., Green, P.R., Leskiar, D.D., Naidu, A., Moffat, B. and Benson, B.J. (1991) Lung surfactant protein SP-C from human, bovine, and canine sources contains plamityl cystine thioester linkages.Am. J. Physiol. 261, 118–25.

    PubMed  Google Scholar 

  • Velander, W.H., Johnson, J.L., Page, R.L., Russell, C.G., Subramanian, A., Wilkins, T.D., Gwazdauskas, F.C., Pittius, C. and Drohan, W.N. (1992a) 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, 12003–7.

    PubMed  Google Scholar 

  • Velander, W.H., Page, R.L., Morcol, T., Russell, C.G., Canesco, R., Young, J.M., Drohan, W.N., Gwazdauskas, F.C., Wilkins, T.D. and Johnson, J.L. (1992b) Production of biologically active human protein C in the milk of transgenic mice.Annals of NY Acad. of Sci. 665, 391–403.

    Google Scholar 

  • Vilotte, J.L., Soulier, S., Stinnarkre, M.G., Massoud, M. and Mercier, J.C. (1989) Efficient expression of bovine α-lactalbumin in transgenic mice.Eur. J. Biochem. 186, 43–8.

    Article  PubMed  Google Scholar 

  • Weaver, T.E. and Whitsett, J.A. (1991) Function and regulation of expression of pulmonary surfactant-associated proteins.Biochem. J. 273, 249–64.

    PubMed  Google Scholar 

  • Wikenheiser, K.A., Vorbroker, D.K., Rice, W.R., Clark, J.C., Bachurski, C.J., Oie, H.K. and Whitsett, J.A. (1993) Production of immortal distal respiratory epithelial lines from surfactant protein C/simian virus 40 large tumor antigen transgenic mice.Proc. Natl. Acad. Sci. USA 90, 11029–33.

    PubMed  Google Scholar 

  • Wright, G., Carver, A., Reeves, D.C.D., Scott, A., Simons, P., Wilmut, I., Garner, I. and Colman, A. (1991) High level expression of active human alpha-1-antitrypsin in the milk of transgenic sheep.Bio/Technology 9, 830–4.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cell Biology, Baylor College of Medicine, One Baylor Plaza, 77030, Houston, TX, USA

    Yongli Wei, Sinai Yarus, Norman M. Greenberg & Jeffrey M. Rosen

  2. Scott Department of Urology, Baylor College of Medicine, One Baylor Plaza, 77030, Houston, TX, USA

    Norman M. Greenberg

  3. Division of Neonatology and Pulmonary Biology, University of Cincinnati College of Medicine and Children's Hospital Medical Center, 45267-0541, Cincinnati, OH, USA

    Jeffrey Whitsett

Authors
  1. Yongli Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sinai Yarus
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Norman M. Greenberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jeffrey Whitsett
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jeffrey M. Rosen
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wei, Y., Yarus, S., Greenberg, N.M. et al. Production of human surfactant protein C in milk of transgenic mice. Transgenic Research 4, 232–240 (1995). https://doi.org/10.1007/BF01969116

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01969116

Keywords

Search

Navigation