Abstract
Owing to its inherent antimicrobial effect and positive charge, the expression of human lysozyme in bovine milk could be beneficial by altering the overal microbial level and the functional and physical properties of the milk. We have used transgenic mice as model systems to evaluate the expression of human lysozyme containing fusion gene constructs in the mammary gland. Expression of human lysozyme was targeted to the mammary gland by using the 5′ promoter elements of either the bovine β (line B mice) or αs1 (line H mice) casein genes coupled to the cDNA for human lysozyme. Expression of human lysozyme mRNA was not found in mammary tissue from any of line B mice. Tissues were analysed from six lines of H mice and two, H6 and H5, were found to express human lysozyme mRNA in the mammary gland at 42% and 116%, respectively, of the levels of the endogenous mouse whey acidic protein gene. At peak lactation, female mice homozygous for the H5 and H6 transgene have approximately twice the amount of mRNA encoding human lysozyme as hemizygous animals. Expression levels of human lysozyme mRNA in the mammary gland at time points representing late pregnancy, early, peak and late lactation corresponded to the profile of casein gene expression. Human lysozyme mRNA expression was not observed in transgenic males, virgin females or in the kidney, liver, spleen or brain of lactating females. A very low level of expression of human lysozyme mRNA was observed in the salivary gland of line H5.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bremel, R.D., Yom, H.C., and Bleck, G.T. (1989) Alteration of milk composition using molecular genetics.J. Diary Sci. 72, 2826–33.
Carlsson, A., Bjorck, L. and Persson, K. (1989) Lactoferrin and lysozyme in milk during acute mastitis and their inhibitory effect in Delvotest P.J. Dairy Sci. 72, 3166–75.
Castanon, M.J., Spevak, W., Adolf, G.R., Chlebowicz-Sledziewska, E. and Sledziewski, A. (1988) Cloning of human lysozyme gene and expression in the yeastSaccharomyces cerevisiae.Gene 66, 223–34.
Chandan, R.C., Parry, R.M. and Shahani, K.M. (1968) Lysozyme, lipase and ribonuclease in milk of various species.J. Dairy Sci. 51, 606–7.
Chang, S.S. (1990) Antimicrobial proteins of maternal and cord sera and human milk in relation to maternal nutritional studies.Am. J. Clin. Nutr. 51, 183–7.
Chomczynski, P. and Sacchi, N. (1987) Single step method of RNA isolation by acid guanidinium thiocyanate phenol chloroform extraction.Anal. Biochem. 162, 156–9.
Clark, A.J., Simons, J.P., Wilmut, I., and Lathe, R. (1987) Pharmaceuticals from transgenic livestock.Trends Biotech. 5, 20–4.
Ebert, K.M., Selgrath, J.P., DiTullio, P., Denman, J., Smith, T.E., Memon, M.A., Schindler, J.E., Monastersky, G.M., Vitale, J.A. and Gorden, K. (1991) Transgenic production of a variant of human tissue type plasminogen activator in goat milk: generation of transgenic goats and analysis of expression.Biol/Technology 9, 835–8.
Eklund, J. and Bradford, G.E. (1976) A note on short term selection for birth weight in mice.Anim. Prod. 22, 127.
Evans, G.A. and Chen, S. (1990) A simple method for screening transgenic mice.Bio Techniques 8, 32–3.
Gorden, K., Lee, E., Vitale, J.A., Smith, A.E., Westphal, H. and Hennighausen, L. (1987) Production of human tissue plasminogen activator in transgenic mouse milk.Bio/ Technology 5, 1183–7.
Gorodetsky, S.I., Tkach, T.M. and Kapelinskaya, T.V. (1988) Isolation and characterization of theBos taurus β-casein gene.Gene 66, 87–96.
Green, M.L. and Marshall, R.J. (1977) The acceleration of cationic materials of the coagulation of casein micelles by rennet.J. Dairy Res. 44, 521–31.
Grinde, B. (1989) A lysozyme isolated from rainbow trout acts on mastitis pathogens.FEMS Micro. Lett. 60, 179–82.
Harris, S., McClenaghan, M., Simons, J.P., Ali, S. and Clark, A.J. (1991) Developmental regulation of the sheep β-lactoglobulin gene in the mammary gland of transgenic mice.Dev. Genet. 12, 299–307.
Hughey, V.L. and Johnson, E.A. (1987) Antimicrobial activity of lysozyme against bacteria involved in food spoilage and food-borne disease.Appl. Environ. Microbiol. 53, 2165–70.
Hughey, V.L., Wilger, P.A. and Johnson, E.A. (1989) Antibacterial activity of hen egg white lysozyme againstListeria monocytogenes ScottA in foods.Appl. Environ. Microbiol. 53(3), 631–8.
Jiminez-Flores, R. and Richardson, T. (1988) Genetic engineering of the caseins to modify behavior of milk processing: a review.J. Dairy Sci. 71, 2640–54.
Jolles, P. and Jolles, J. (1984) What's new in lysozyme research.Mol. Cell. Biochem. 63, 165–89.
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.
Maga, E.A. and Richardson, T. (1991) Amplification of a 9.0 kb fragment using PCR.Bio/Techniques 11, 185–6.
Meade, H., Gates, L., Lacy, 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.
Panfil-Kuncewicz, H. and Kisza, J. (1976) Studies on the lysozyme content of human and cow milk.Zesz. Nauk. Akad. Roln.-Tech. Olsztynie, Technol. Zywn. 8, 105–12.
Phillips, D.C. (1966) The 3-dimensional structure of an enzyme molecule.Sci Am. 215, 78–90.
Redhead, K., Hill, T. and Mulloy, B. (1990) Antimicrobial effect of human milk onBordetella pertussis.FEMS Micro. Lett. 70, 269–74.
Shani, M., Barash, I., Nathan, M., Ricca, G., Searfoss, G.H., Dekel, I., Faerman, A., Givol, D. and Hurwitz, D.R. (1992) Expression of human serum albumin in the milk of transgenic mice.Transgenic Res. 1, 195–208.
Simons, J.P., McClenaghan, M. and Clark, A.J. (1987) Alteration of the quality of milk by expression of β-lactoglobulin in transgenic mice.Nature 328, 530–2.
Vilotte, J.L., Solulier, S., Stinnakre, M.G., Massoud, M. and Mercier, J.C. (1989) Efficient tissue specific expression of bovine α-lactalbumin in transgenic mice.Eur. J. Biochem. 186, 43–8.
Whitelaw, C.B.A., Archibald, A.L., Harris, S., McClenaghan, M., Simons, J.P. and Clark, A.J. (1991) Targeting expression to the mammary gland: intronic sequences can enhance the efficiency of gene expression in transgenic mice.Transgenic Res. 1, 3–13.
Wilmut, I., Archibald, A.L., McClenaghan, M., Simons, J.P., Whitelaw, C.B.A. and Clark, A.J. (1991) Production of pharmaceutical proteins in milk.Experientia 47, 905–12.
Wright, G., Carver, A., Cottom, D., Reeves, D., Scott, A., Simons, P., Wilmut, I., Garner, I. and Colman, A. (1991) High level expression of active human α-1-antitrypsin in the milk of transgenic sheep.Bio/Technology 9, 830–4.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Maga, E.A., Anderson, G.B., Huang, M.C. et al. Expression of human lysozyme mRNA in the mammary gland of transgenic mice. Transgenic Research 3, 36–42 (1994). https://doi.org/10.1007/BF01976025
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01976025