Murine α-Lactalbumin Gene Inactivation and Replacment

  • Alexander Stacey
  • Angelika Schnieke


We are using gene targeting to investigate the role of α-lactalbumin in lactation and the regulation of α-lactalbumin gene expression. Gene targeting in embryonic stem (ES) cells is now widely used to inactivate genes of interest. We have recently demonstrated a powerful technique, termed double replacement, whereby a wide variety of genetic alterations can be engineered at a chosen locus and the effects examined in mice (Stacey et al.,1994) The procedure involves two consecutive rounds of gene targeting: the first creates a null allele of the gene of interest, which can then be replaced with DNA carrying experimental alterations in the second round. This process effects a clean replacement of DNA sequences, and no unwanted exogenous DNA (eg marker genes) are left at the target site. A most useful aspect of double replacement gene targeting is that multiple independent second step targeting experiments can be carried out, making it possible to produce and compare many experimental alterations at the same locus.


Embryonic Stem Cell Mammary Gland Embryonic Stem Cell Line Consecutive Round Experimental Alteration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. Fitzgerald, D.K., Brodbeck, U., Kiyosawa, I., Mawal, R., Colvin, B. and Ebner, K.E., 1970, α- lactalbumin and the lactose synthetase reaction, J. Biol. Chem. 245:2103.PubMedGoogle Scholar
  2. Magin, T.M., McWhir, J. and Melton, D.W., 1992, A new mouse embryonic stem cell line with good germ line contribution and gene targeting frequency, Nucl. Acids Res. 14:3795.CrossRefGoogle Scholar
  3. Schmidt, D.V., Walker, I.E. and Ebner, K.E., 1971, Lactose synthetase activity in northern fur seal milk, Biochim. et Biophys. Acta 252:439.CrossRefGoogle Scholar
  4. Soulier, S., Vilotte, J.L., Stinnakre, M.G, and Mercier, J.C., 1992, Expression analysis of ruminant α- lactalbumin in transgenic mice: developmental regulation and general location of important cis- regulatory elements, F.E.B.S. Letts. 297:13.CrossRefGoogle Scholar
  5. Stacey, A.J., Schnieke, A., McWhir, J., Cooper, J., Colman, A. and Melton, D.W., 1994, Use of double replacement gene targeting to replace the murine α-lactalbumin gene with its human counterpart in embryonic stem cells and mice, Mol. Cell Biol. 14:1009.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Alexander Stacey
    • 1
  • Angelika Schnieke
    • 1
  1. 1.Pharmaceutical Proteins Ltd.RoslinUK

Personalised recommendations