Abstract
The main nutritional limitation of maize used for feed is the content of protein that is digestible, bioavailable and contains an amino acid balance that matches the requirements of animals. In contrast, milk protein has good digestibility, bioavailability and amino acid balance. As an initial effort to create maize optimized as a source of swine nutrition, a codon-adjusted version of a gene encoding the milk protein porcine α-lactalbumin was synthesized. Maize expression vectors containing this gene under the control of the Ubi-1 promoter and nos 3′ terminator were constructed. These vectors were used to transform maize callus lines that were regenerated into fertile plants. The α-lactalbumin transgenes were transmitted through meiosis to the sexual progeny of the regenerated plants. Porcine α-lactalbumin was detected in callus and kernels from transgenic maize lines that were transformed by two constructs containing the 27-kDa maize gamma-zein signal sequence at the 5′ end of the synthetic porcine α-lactalbumin coding sequence. One of these constructs contained an ER retention signal and the other did not. Expression was not observed in kernels or callus from transgenic maize lines that were transformed by a construct that does not contain an exogenous protein-targeting signal. This suggests that the signal peptide might play an important role in porcine α-lactalbumin accumulation in transgenic maize kernels.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Altenbach SB, Kuo C-C, Staraci LC, Pearson KW, Wainwright C, Georgescu A et al. (1992) Accumulation of a brazil nut albumin in seeds of transgenic canola results in enhances levels of seed protein methionine. Plant Mol Biol 18: 235–245.
Armstrong CL, Green CE and Phillips RL (1991) Development and availability of germplasm with high type II culture formation response. Maize Genet Coop Newsletter 65: 92–93.
Adang MJ, Brody MS, Cardineau G, Eagan N, Roush RT, Shewmaker CK et al. (1993) The reconstruction and expression of a Bacillus thuringiensis cryIIIA gene in protoplast and potato plants. Plant Mol Biol 21: 1131–1145.
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254.
Brar DS, Ohtani T and Uchimiya H (1995) Genetically engineered plants for quality improvement. Biotech Genet Eng Rev 13: 167–179.
Chaïr H, Legavre T and Guiderdoni E (1996) Transformation of haploid, microspore-derived cell suspension protoplasts of rice (Oryza sativa L.). Plant Cell Rep 15: 766–770.
Christensen AH, Sharrock RA and Quail PH (1992) Maize polyubiquitin genes: structure thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Mol Biol 18: 675–689.
Chong DKX, Roberts W, Arakawa T, Illes K, Bagi G, Slattery CW et al. (1997) Expression of the human milk protein β-casein in transgenic potato plants. Transgenic Res 6: 289–296.
Cornejo M-J, Luth D, Blechl AE and Anderson OD (1993) Activity of a maize ubiquitin promoter in transgenic rice (abstract No. 109). Plant Physiol 99: S-19.
Das Gupta NA, Alexander LJ and Beattie CW (1992) The sequence of a porcine cDNA encoding α-lactalbumin. Gene 110: 265–266.
Fiedler U and Conrad U (1995) High-level production and long-term storage of engineered antibodies in transgenic tobacco seeds. Bio/Technology 13: 1090–1093.
Frame BR, Zhang H, Cocciolone S, Sidorenko LV, Dietrich CR, Pegg SE et al. (2000) Production of transgenic maize from bombarded type II callus: Effect of gold particle size and callus morphology on transformation efficiency. In Vitro Cell Dev Biol-Plant 36: 21–29.
Harlow E and Lane D (1988) Antibodies, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Hood EE, Witcher DR, Maddock S, Meyer T, Baszczynski C, Bailey M et al. (1997) Commercial production of avidin from transgenic maize: characterization of transformants, production, processing, extraction and purification. Mol Breeding 3: 291–306.
Horvath H, Huang J, Wong O, Kohl E, Okita T, Kannangara CG et al. (2000) The production of recombinant proteins in transgenic barley grains Proc Natl Acad Sci USA 97: 1914–1919.
Kitamura K (1993) Breeding trials for improving the food-processing quality of soybeans. Trends Food Sci Technol 4: 64–67.
Klee WA and Klee CB (1970) The role of α-lactalbumin in lactose synthesis. Biochem Biophys Res Commun 39: 833–841.
Koziel MG, Beland GL, Bowman C, Carozzi NB and Crenshaw R (1993) Field performance of elite transgenic maize plants expressing an insecticidal protein derived from Bacillus thuringiensis. Bio/Technology 11: 194–200.
Kusnadi AR, Nikolov ZA and Howard JA (1997) Production of recombinant proteins in transgenic plants: Practical considerations. Biotech and Bioeng 5: 473–484.
Leammli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227: 680–685.
Mitra A and Zhang Z (1994) Expression of a human lactoferrin cDNA in tobacco cells produces antibacterial protein(s). Plant Physiol 106: 977–981.
Molvig L, Tabe LM, Eggum BO, Moore AE, Craig S, Spencer D et al. (1997) Enhanced methionine levels and increased nutritive value of seeds of transgenic lupins (Lupinus angustifolius L.) expressing a sunflower seed albumin gene. Proc Natl Acad Sci USA 94: 8393–8398.
Nayak P, Basu D, Das S, Basu A, Ghosh D, Ramakrishnan NA et al. (1997) Transgenic elite indica rice plants expressing CryIAc δ-endotoxin of Bacillus thuringiensis are resistant against yellow stem borer (Scirpophaga incertulas). Proc Natl Acad Sci USA 94: 2111–2116.
Payne PI (1983) Breeding for protein quantity and protein quality in seed crops. In: Daussant J, Mossé J and Vaughan J (eds), Annual Proceedings of the Phytochemical Society of Europe No. 20, Seed Proteins pp. 223–253. Academic Press, London.
Pen J, Sijmons PC, van Ooijen AJJ and Hoekema A (1993) Protein production in transgenic crops: analysis of plant molecular farming. In: Hiatt A (ed), Transgenic Plants: Fundamentals and Applications. pp. 239–251 Marcel Dekker, New York.
Perlak FJ, Fuchs RL, Dean DA, Mcpherson SL and Fischhoff DA (1991) Modification of the coding sequence enhances plant expression of insect control protein genes. Proc Natl Acad Sci USA 88: 3324–3328.
Prat S, Cortadas J, Puigdomenech P, and Palau J (1985) Nucleic acid (cDNA) and amino acid sequences of the maize endosperm protein glutelin-2. Nucleic Acids Res 13: 1493–1504.
Robinson, RK (1986) Modern Dairy Technology, vol. 1. Advances in Milk Processing. Elsevier Applied Science Publishers, New York.
Rouwendal GJA, Mendes O, Walbert EJH and Boer DD (1997) Enhanced expression in tobacco of the gene encoding green fluorescent protein by modification of its codon usage. Plant Mol Biol 33: 989–999.
Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, a Laboratory Mannual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.
Sardana R, Dukiandjiev S, Giband M, Cheng X, Cowan K, Sauder C et al. (1996) Construction and rapid testing of synthetic and modified toxin gene sequences CryIA (b&c) by expression in maize endosperm culture. Plant Cell Rep. 15: 677–681.
Schouten A, Roosien J, van Engelen FA, deJong GAM, Borst-Vrenssen AWM, Zilverentant JF et al. (1996) The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be both the cytosol and the secretory pathway in transgenic tobacco. Plant Mol Biol 30: 781–793.
Sijmons PC, Dekker BMM, Schrammeijer B, Verwoerd TC, van den Elzen PJM and Hoekema A (1990) Production of correctly processed human serum albumin in transgenic plants. Bio/Technology 8: 217–221.
Stewart CJ, Adang MJ, All JN, Raymer PL, Ramachandran S and Parrot WA (1996) Insect control and dosage effects in transgenic canola containing a synthetic Bacillus thuringiensis cryIAc gene. Plant Physiol 112: 115–120.
Takase K and Hagiwara K (1998) Expression of human α-lactalbumin in transgenic tobacco. J Bioche m 123: 440–444.
Taylor MG, Vasil V and Vasil IK (1993) Enhanced GUS gene expression in cereal/grass cell suspensions and immature embryos using the maize ubiquitin-based plasmid pAHC25. Plant Cell Rep 12: 491–495.
Toki S, Takamatsu S, Norjiri C, Ooba S, Anzai H, Iwata M et al. (1992) Expression of a maize ubiquitin gene promoter-bar chimeric gene in transgenic rice plants. Plant Physiol 100: 1503–1507.
Wandelt CI, Khan MRI, Craig S, Schroeder HE, Spencer D and Higgins TJV (1992) Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants. Plant J 2: 181–192.
Weeks JT, Anderson OD and Blechl AE (1993) Rapid production of multiple independent lines of fertile transgenic wheat (Triticum aestivum). Plant Physiol 102: 1077–1084.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Yang, SH., Moran, D.L., Jia, HW. et al. Expression of a Synthetic Porcine α-Lactalbumin Gene in the Kernels of Transgenic Maize. Transgenic Res 11, 11–20 (2002). https://doi.org/10.1023/A:1013996129125
Issue Date:
DOI: https://doi.org/10.1023/A:1013996129125