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Proc Natl Acad Sci U S A. Jul 5, 1994; 91(14): 6544–6548.
PMCID: PMC44239

Creation and phenotypic analysis of alpha-lactalbumin-deficient mice.

Abstract

alpha-Lactalbumin is an abundant milk-specific calcium metalloprotein which has an evolutionary relationship to lysozyme. It modifies the substrate specificity of a Golgi galactosyltransferase by forming the lactose synthetase binary complex. Lactose, together with other sugars and diffusible ions, is responsible for the osmotic pressure of milk. To assess the involvement of alpha-lactalbumin in lactogenesis, alpha-lactalbumin-deficient mice were created by disrupting the gene by homologous recombination in embryonic stem cells. Homozygous mutant mice are viable and fertile but females cannot feed their offspring. They produce a highly viscous milk that pups appear to be unable to remove from the mammary gland. This milk is rich in fat and protein and is devoid of alpha-lactalbumin and lactose. The phenotype of heterozygous mice was found to be intermediate, with a 40% decrease in alpha-lactalbumin but only a 10-20% decrease in the lactose content of their milk compared with wild-type animals. These results emphasize the key function of alpha-lactalbumin in lactogenesis and open new opportunities to manipulate milk composition.

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  • Ebner K, Brodbeck U. Biological role of alpha-lactalbumin: a review. J Dairy Sci. 1968 Mar;51(3):317–322. [PubMed]
  • Pilson ME, Kelly AL. Composition of the Milk from Zalophus californianus, the California Sea Lion. Science. 1962 Jan 12;135(3498):104–105. [PubMed]
  • Messer M, Elliott C. Changes in alpha-lactalbumin, total lactose, UDP-galactose hydrolase and other factors in tammar wallaby (Macropus eugenii) milk during lactation. Aust J Biol Sci. 1987;40(1):37–46. [PubMed]
  • Komine S, Nakanishi K, Anzai T, Yoshimoto A. Process of the combination of alpha-lactalbumin with lysosomes in mammary epithelial cells. Cell Biochem Funct. 1985 Jan;3(1):41–44. [PubMed]
  • Komine S, Nakanishi K, Anzai T, Yoshimoto A. Alpha-lactalbumin as a lysosomal enzyme-releasing factor. Cell Biochem Funct. 1985 Jan;3(1):33–39. [PubMed]
  • Vilotte JL, Soulier S. Isolation and characterization of the mouse alpha-lactalbumin-encoding gene: interspecies comparison, tissue- and stage-specific expression. Gene. 1992 Oct 1;119(2):287–292. [PubMed]
  • Lathe R, Vilotte JL, Clark AJ. Plasmid and bacteriophage vectors for excision of intact inserts. Gene. 1987;57(2-3):193–201. [PubMed]
  • Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R. The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium. J Embryol Exp Morphol. 1985 Jun;87:27–45. [PubMed]
  • Vilotte JL, Soulier S, Stinnakre MG, Massoud M, Mercier JC. Efficient tissue-specific expression of bovine alpha-lactalbumin in transgenic mice. Eur J Biochem. 1989 Dec 8;186(1-2):43–48. [PubMed]
  • Soulier S, Vilotte JL, Stinnakre MG, Mercier JC. Expression analysis of ruminant alpha-lactalbumin in transgenic mice: developmental regulation and general location of important cis-regulatory elements. FEBS Lett. 1992 Feb 3;297(1-2):13–18. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • Mansour SL, Thomas KR, Capecchi MR. Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature. 1988 Nov 24;336(6197):348–352. [PubMed]
  • te Riele H, Maandag ER, Berns A. Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5128–5132. [PMC free article] [PubMed]
  • van Deursen J, Wieringa B. Targeting of the creatine kinase M gene in embryonic stem cells using isogenic and nonisogenic vectors. Nucleic Acids Res. 1992 Aug 11;20(15):3815–3820. [PMC free article] [PubMed]
  • Stacey A, Schnieke A, McWhir J, Cooper J, Colman A, Melton DW. Use of double-replacement gene targeting to replace the murine alpha-lactalbumin gene with its human counterpart in embryonic stem cells and mice. Mol Cell Biol. 1994 Feb;14(2):1009–1016. [PMC free article] [PubMed]
  • Shulman MJ, Nissen L, Collins C. Homologous recombination in hybridoma cells: dependence on time and fragment length. Mol Cell Biol. 1990 Sep;10(9):4466–4472. [PMC free article] [PubMed]
  • Hasty P, Rivera-Pérez J, Bradley A. The length of homology required for gene targeting in embryonic stem cells. Mol Cell Biol. 1991 Nov;11(11):5586–5591. [PMC free article] [PubMed]
  • Yagi T, Ikawa Y, Yoshida K, Shigetani Y, Takeda N, Mabuchi I, Yamamoto T, Aizawa S. Homologous recombination at c-fyn locus of mouse embryonic stem cells with use of diphtheria toxin A-fragment gene in negative selection. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9918–9922. [PMC free article] [PubMed]
  • McCarrick JW, 3rd, Parnes JR, Seong RH, Solter D, Knowles BB. Positive-negative selection gene targeting with the diphtheria toxin A-chain gene in mouse embryonic stem cells. Transgenic Res. 1993 Jul;2(4):183–190. [PubMed]
  • Mombaerts P, Iacomini J, Johnson RS, Herrup K, Tonegawa S, Papaioannou VE. RAG-1-deficient mice have no mature B and T lymphocytes. Cell. 1992 Mar 6;68(5):869–877. [PubMed]
  • Zijlstra M, Li E, Sajjadi F, Subramani S, Jaenisch R. Germ-line transmission of a disrupted beta 2-microglobulin gene produced by homologous recombination in embryonic stem cells. Nature. 1989 Nov 23;342(6248):435–438. [PubMed]
  • Rennison ME, Kerr M, Addey CV, Handel SE, Turner MD, Wilde CJ, Burgoyne RD. Inhibition of constitutive protein secretion from lactating mouse mammary epithelial cells by FIL (feedback inhibitor of lactation), a secreted milk protein. J Cell Sci. 1993 Oct;106(Pt 2):641–648. [PubMed]

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