Summary
Owing to its outstanding high and avid Fe3+ ion-binding capacity, the secreted 80-kDa glycoprotein lactoferrin (Lf) is thought to act as an antimicrobial agent against a broad spectrum of microbial species. To examine the proposed role of the bovine Lf-encoding gene as a candidate gene for conferring resistance against microbial infections, we systematically searched for natural variants of the gene and its promoter. Based on our previous characterization of the bovine Lf-encoding gene and published cDNA sequences, we identified 36 potentially polymorphic sites within the coding region of the gene, 15 of them altering the protein sequence. Experimental analysis confirmed two of these polymorphisms from among 11 sites checked in a variety of bovine breeds. These exchanges are tightly linked on two alleles. Comparative sequencing in dairy and beef breeds revealed 17 polymorphic sites within approx 700 bp of the proximal promoter region analyzed. One of these was proven to result in a polymorphic SP1-box. The promoter analysis within these breeds and in related Bovidae species suggests that the relatively low expression of the bovine Lf-gene is not a consequence of domestication.
In order to establish a model system for an experimental examination of the Lf resistance capacity against microbial infections, we inserted a bovine Lf cDNA copy into regulatory segments of the bovine αs1-casein gene, for hyperexpression in the mammary gland. Western blotting confirmed its expression in a murine MEC cell line. Genomic transfer of the fusion gene gave rise to two male mice having stably incorporated the transgene into their genome, and are now being used as founders to establish transgenic lines.
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References
Anderson, B. F., Baker, H. M., Dodson, E. J., Norris, G. E., Rumball, S. V., Waters, J. M., and Baker, E. N. (1987) Structure of the human lactoferrin at 3.2 A resolution. Proc. Natl Acad. Sci. USA 84, 1769–1773.
Anderson, B. F., Baker, H. M, Norris, G. E., Rice, D. W., and Baker, E. N. (1989) Structure of the human lactoferrin: crystallographic structure analysis and refinement at 2.8A resolution. J. Mol. Biol. 209, 711–734.
Anderson, B. F., Baker, H. M., Norris, G. E., Rumball, S. V., and Baker, N. (1990) Apolactoferrin structure demonstrates ligand-induced conformational change in transferrins. Nature 344, 784–787.
Ball, R. K., Friis, R. R., Schoenenberger, C. A., Doppler, W., and Groner, B. (1988) Prolactin regulation of ß-casein gene expression and of a cytosolic 120-kd protein in a cloned mouse mammary epithelial cell line. EMBO J. 7, 2089–2095.
Birgens, H. S. (1991) The interaction of lactoferrin with human monocytes. Dan. Med. Bull. 38, 244–252.
Bullen, J. J., Rogers, H. J., and Leigh, L. (1972) Iron-binding proteins in milk and resistance to Escherichia coli infections in infants. Br. Med. J. 1, 69–75.
Cramer, E., Pryzwansky, K. B., Villeval, J. L., Testa, U., and Brenton-Gorius, J. (1985) Ultrastructural localization of lactoferrin and myeloperoxidase in human neutrophiles by immunogold. Blood 65, 423–432.
Don, R. H., Cox, P. T., Wainwright, B. J., Baker, K., and Mattick, J. S. (1991) “Touchdown” PCR to circumvent spurious priming during gene amplification. Nucleic Acids Res. 19, 4008.
Erhardt, G., Meyer, F., and Senft, B. (1982) Umweltbedingte und genetische Aspekte der Mastitis. Züchtungskunde 54, 86–105.
Furmanski, P, Li, Z.-P, Fortuna, M. B., Swamy, Ch.V. B., and Das, R. (1989) Multiple molecular forms of human lactoferrin. J. Exp. Med. 170, 415–429.
Gaunt, S. N., Raffio, N., Kingsbury, E. T., Damon, R. A., Jr., Johnson, W. H., and Mitchell, B. A. (1980) Variation of lactoferrin and mastitis and their heritabilities. J. Dairy Sci. 63, 1874–1880.
Goodman, R. E. and Schanbacher, F. L. (1991) Bovine lactoferrin mRNA: sequence, analysis and expression in mammary gland. Biophys. Res. Commun. 180, 75–84.
Hogan, B., Costantini, F., and Lacy, E. (1986) Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Johnston, J. J., Rintels, P., Chung, J., Sather, J., Benz, E. J., Jr., and Berliner, N. (1992) Lactoferrin gene promoter: structural integrity and nonexpression in HL60 cells. Blood 79, 2998–3006.
Koczan, D. (1994) Das bovine asl-Casein Gen: Sequenz, Allelvarianten, Expressionsregulation und Biotechnologische Anwendung. Thesis, University of Gießen, FRG.
Koczan, D., Hobom, G., and Seyfert, H.-M. (1991) Genomic organization of the bovine alphaSl casein gene. Nucleic Acids Res. 19(20), 5591–5596.
Liu, Y. and Teng, C. T. (1991) Characterization of estrogen-responsive mouse lactoferrin promoter. J. Biol. Chem. 266, 21,880–21,885.
Marzurier, J. and Spik, G. (1980) Comparative study of the iron-binding properties of human transferrins. I. Complete and sequential iron saturation and desaturation of the lactotransferrin. Biochim. Biophys. Acta 629, 399–406.
Masson, P. L. and Heremans, J. F. (1971) Lactoferrin in milk from different species. Comp. Biochem. Physiol 39B, 119–129.
Mead, P. E. and Tweedie, J. W. (1990) cDNA and protein sequence of bovine lactoferrin. Nucleic Acids Res. 18, 7167.
Natour, S. and Seyfert, H.-M. (1994) Quantitative control of casein synthesis: delineation of an α-s1-CN RNA sequence motif conferring hormonal mRNA stabilization in cattle. XXIV International Conference on Animal Genetics, Prague, E67 (abstract).
Pierce, A., Colavizza, D., Benaissa, M., Maes, P., Tartar, A., and Montreuil, J. (1991) Molecular cloning and sequence analysis of bovine lactotransferrin. Eur. J. Biochem. 196, 177–184.
Sanchez, L., Calvo, M., and Brock, J. H. (1992) Biological role of lactoferrin. Arch. Dis. Child. 67, 657–661.
Sawatzki, G. (1987) The role of iron binding proteins in bacterial infections, in Iron Transport in Microbes, Plants and Animals (Winkelmann, A., von der Helm, P., and Neilands, U., eds.), VCH, Weinheim, pp. 477–498.
Schanbacher, F. L., Goodman, R. E., and Talhouk, R. S. (1993) Bovine mammary lactoferrin: implications from messenger ribonucleic acid (mRNA) sequence and regulation contrary to other milk proteins. J. Dairy Sci. 76, 3812–3831.
Schutz, M. M. (1994) Genetic evaluation of somatic cell scores for United States dairy cattle. J. Dairy Sci. 77, 2113–2129.
Seyfert, H.-M. and Kühn, C. (1994) Characterization of a first bovine lactoferrin gene variant, based on an EcoRI polymorphism. Anim. Genet. 25, 54.
Seyfert, H.-M. and Sawatzki, G. (1986) An estimation of the soluble tubulin content in Tetrahymena cells of normal and of size altered phenotype. Exp. Cell Res. 162, 86–96.
Seyfert, H.-M., Tuckoricz, A., Interthal, H., Koczan, D., and Hobom, G. (1994) Structure of the bovine lactoferrin-encoding gene and its promoter. Gene 143, 265–269.
Simianer, H., Solbu, H., and Schaeffer, L. R. (1991) Estimated genetic correlations between disease and yield traits in dairy cattle. J. Dairy Sci. 74, 4358–4362.
Southern, P. J. and Berg, P. (1982) Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J. Mol. Appl Genet. 1, 327–341.
Stewart, A. F., Willis, J. M., and Mackinlay, A. G. (1984) Nucleotide sequences of bovine aSl and K-casein cDNAs. Nucleic Acids Res. 12, 3895–3907.
Tomita, M. W., Bellamy, M., Takase, K., Yamauchi, H., Wakabayashi, H., and Kawase, K. (1991) Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. J. Dairy Sci. 74, 4137–4142.
Tsuji, S., Hirata, Y., and Matsuoka, K. (1989) Two apparaent molecular forms of bovine lactoferrin. J. Dairy Sci. 72, 1130–1136.
Wall, D. A., David, S. K., and Read, B. M. (1992) Phylogenetic relationship in the subfamily Bovinae (Mammalia: Artiodactyla) based on ribosomal DNA. J. Mammology 73, 262–275.
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Seyfert, HM., Klußmann, U., Steinhoff, U.M., Vanselow, J., Koczan, D., Hobom, G. (1997). Variants and Biotechnological Use of the Bovine Lactoferrin-Encoding Gene. In: Hutchens, T.W., Lönnerdal, B. (eds) Lactoferrin. Experimental Biology and Medicine, vol 28. Humana Press. https://doi.org/10.1007/978-1-4612-3956-7_4
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DOI: https://doi.org/10.1007/978-1-4612-3956-7_4
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