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Development of transgenic rice lines expressing the human lactoferrin gene

  • Lee, Jin-Hyoung (National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Il-Gi (Department of Biomedical Science, Hallym University) ;
  • Kim, Hyo-Sung (National Academy of Agricultural Science, Rural Development Administration) ;
  • Shin, Kong-Sik (National Academy of Agricultural Science, Rural Development Administration) ;
  • Suh, Seok-Cheol (National Academy of Agricultural Science, Rural Development Administration) ;
  • Kweon, Soon-Jong (National Academy of Agricultural Science, Rural Development Administration) ;
  • Rhim, Seong-Lyul (Department of Biomedical Science, Hallym University)
  • Received : 2010.11.15
  • Accepted : 2010.11.29
  • Published : 2010.12.31

Abstract

Lactoferrin is an 80-kDa iron-binding glycoprotein that is found in high concentrations in human milk. Human lactoferrin (hLF) has several beneficial biological activities including immune system modulation and antimicrobial activity. In the present study, we devolved a method of hLF expression through introducing the hLF gene construct into Oriza sativa cv. Nakdong using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the rice glutelin promoter was detected in the seeds of transgenic rice plants. Transformed rice plants were selected on media containing herbicide(DL-phosphinothricin) and the integration of hLF cDNA was confirmed by Southern blot analysis. The expression of the full length hLF protein from the grains of transgenic rice plants was verified by Western blot analysis. The lactoferrin expression levels in the transformed rice grains determined by enzyme-linked immunosorbant assay accounted for approximately 1.5% of total soluble protein. Taken together, these data indicate that rice grains expressing hLF can be directly incorporated into infant formula and baby food.

Keywords

References

  1. Arakawa T, Chong DKX, Slattery CW, Langridge WHR (1999) Improvements in human health through production of human milk proteins in transgenic food plants. In: Shahidi F, Kolodziejczyk P, Whitaker JR (eds) Chemicals via Higher Plant Bioengineering, Kluwer Academic/Plenum Publishers, New York: 149-159
  2. Bradford M (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 https://doi.org/10.1016/0003-2697(76)90527-3
  3. Choi SM, Lee OS, Kwon SY, Kwak SS, Yu DY, Lee HS (2003) High expression of a human lactoferrin in transgenic tobacco cell cultures. Biotech Lett 25:213-218 https://doi.org/10.1023/A:1022341917735
  4. Chong DKX, Langridge WHR (2000) Expression of full-length bioactive antimicrobial human lactoferrin in potato plants. Transgenic Res 9:71-78 https://doi.org/10.1023/A:1008977630179
  5. Chong DKX, Roberts W, Arakawa T, Illes K, Bagi G, Slattery CW, Langridge WHR (1997) Expression of the human milk protein $\beta$-casein in transgenic potato plants. Transgenic Res 6:289-296 https://doi.org/10.1023/A:1018410712288
  6. Chu CC, Wang CC, Sun CS, Hsu C, Yin KC, Chu CY, Bi FY (1975) Establishment of an efficient medium for another culture of rice through comparative experiments on the nitrogen sources. Sci Sin 18:659-668
  7. Hanson LA and Korotkova M (2002) The role of breastfeeding in prevention of neonatal infection, Seminars in fetal & neonatal medicine. 7:275-281 https://doi.org/10.1016/S1084-2756(02)90124-7
  8. Lee TJ, Coyne DP, Clemente TE, Mitra A (2002) Partial resistance to bacterial wilt in transgenic tomato plants expressing antibacterial lactoferrin gene. J Amer Soc Hort Sci 127:158-164
  9. Li Y, Geng Y, Song H, Zheng G, Huan L, Qiu B (2004) Expression of a human lactoferrin N-lobe in Nicotiana benthamiana with potato virus X-based agroinfection. Biotechnol Lett 26:953-957 https://doi.org/10.1023/B:BILE.0000030038.27358.20
  10. Liang Q, Richardson T (1993) Expression and characterization of human lactoferrin in yeast Saccharomyces cerevisiae. J Agric Food Chem 41:1800-1807 https://doi.org/10.1021/jf00034a053
  11. Lonnerdal B (1996) Recombinant human milk proteins-an opportunity and a challenge. Am J Clin Nutr 63:622S-626S
  12. Lonnerdal B (2002) Expression of human milk proteins in plants. J Am Clin Nutr 21:218S-221S https://doi.org/10.1080/07315724.2002.10719269
  13. Min SR, Woo JW, Jeong WJ, Han SK, Lee YB, Liu JR (2006) Production of human lactoferrin in transgenic cell suspension cultures of sweet potato. Biologia Plantarum 50:131-134 https://doi.org/10.1007/s10535-005-0087-5
  14. Murray M and Thopson WF (1980) Rapid isolation of molecular weight plant DNA. Nucleic Acid Res., 8:4321-5 https://doi.org/10.1093/nar/8.19.4321
  15. Nandi S, Suzuki YA, Huang J, Yalda D, Pham P, Wu L, Bartley G, Huang N, Lonnerdal B (2002) Expression of human lactoferrin in transgenic rice grains for the application in infant formula. Plant Sci 163:713-722 https://doi.org/10.1016/S0168-9452(02)00165-6
  16. Nandi S, Yalda D, Lu S, Nikolov Z, Misaki R, Fujiyama K, Huang N (2005) Process development and economic evaluation of recombinant human lactoferrin expressed in rice grain. Transgenic Res 14:237-249 https://doi.org/10.1007/s11248-004-8120-6
  17. Platenburg GJ, Kootwijk EP, Kooiman PM, Woloshuk SL, Nuijens JH, Krimpenfort PJ, Peeper FR, de Boer HA, Strijker R (1994) Expression of human lactoferrin in milk of transgenic mice. Transgenic Res 3:99-108 https://doi.org/10.1007/BF01974087
  18. Philip MG, Bowler C (2002) Molecular plant biology volume one: a practical approach. Oxford University Press, Oxford: UK 258:17-23
  19. Salmon V, Legrand D, Slomianny MC, Yazidi IE, Spik G, Gruber V, Bournat P, Olagnier B, Mison D, Theisen M, Merot B (1998) Production of human lactoferrin in transgenic tobacco plants. Protein Express Purif 13:127-135 https://doi.org/10.1006/prep.1998.0886
  20. Simon R (1984) High frequency mobilization of gram negative bacterial replicons by the in vitro constructed Tn5-Mob transposon. Mol Gen Genet 196:413-420 https://doi.org/10.1007/BF00436188
  21. Van Berkel PHC, Welling MM, Geerts M, Van Veen HA, Ravensbergen B, Salaheddine M, Pauwels EKJ, Pieper F, Nuijens JH, Nibbering PH (2002) Large scale production of recombinant human lactoferrin in the milk of transgenic cows. Nature Biotechnol 20:484-487 https://doi.org/10.1038/nbt0502-484
  22. Ward PP, May GS, Headon DR, Conneely OM (1992) An inducible expression system for the production of human lactoferrin in Aspergillus nidulans. Gene 122:219-223 https://doi.org/10.1016/0378-1119(92)90054-S

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