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Accumulation, assembly, and digestibility of amarantin expressed in transgenic tropical maize

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Abstract

An amaranth (Amaranthus hypochondriacus) 11S globulin cDNA, encoding one of the most important storage proteins (amarantin) of the seed, with a high content of essential amino acids, was used in the transformation of CIMMYT tropical maize genotype. Constructs contained the amarantin cDNA under the control of a tissue-specific promoter from rice glutelin-1 (osGT1) or a constitutive (CaMV 35S) promoter with and without the first maize alcohol dehydrogenase intron (AdH). Southern-blot analysis confirmed the integration of the amarantin cDNA, and copy number ranged from one to more than ten copies per maize genome. Western-blot and ultracentrifugation analyses of transgenic maize indicate that the expressed recombinant amarantin precursors were processed into the mature form, and accumulated stably in maize endosperm. Total protein and some essential amino acids of the best expressing maize augmented 32% and 8–44%, respectively, compared to non-transformed samples. The soluble expressed proteins were susceptible to digestion by simulated gastric and intestinal fluids, and it is suggested that they show no allergenic activity. These findings demonstrate the feasibility of using genetic engineering to improve the amino acid composition of grain crops.

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Acknowledgements

The authors would like to thank Dr. J. Calderon for polyclonal antibody samples and Dr. F. Guevara, from CINVESTAV-IPN, for discussion and reviewing this paper. Thanks are also due to Dr. S. McLean for greenhouse work, Dr. R. J. Peña for analytical services, and to P. Estañol and M. Pacheco for excellent technical assistance, all from CIMMYT. The financial support from CONACYT-México is acknowledged as well.

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Author notes

  1. N. Bohorova

    Present address: Epicyte Pharmaceutical Inc., 5810 Nanacy Ridge Drive, 150, San Diego, CA 92121, USA

Authors and Affiliations

  1. Depto. de Biotecnología y Bioquímica, Centro de Investigación y de Estudios Avanzados del IPN, Apdo. Postal 629, 36500, Irapuato, Gto., México

    Q. Rascón-Cruz, S. Sinagawa-García, J. A. Osuna-Castro & O. Paredes-López

  2. CIMMYT, Apdo. Postal 6–641, 06600, México, DF, México

    N. Bohorova

Authors
  1. Q. Rascón-Cruz
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  2. S. Sinagawa-García
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  3. J. A. Osuna-Castro
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  4. N. Bohorova
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  5. O. Paredes-López
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Correspondence to O. Paredes-López.

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Communicated by J.W. Snape

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Rascón-Cruz, Q., Sinagawa-García, S., Osuna-Castro, J.A. et al. Accumulation, assembly, and digestibility of amarantin expressed in transgenic tropical maize. Theor Appl Genet 108, 335–342 (2004). https://doi.org/10.1007/s00122-003-1430-x

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  • DOI: https://doi.org/10.1007/s00122-003-1430-x

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