Development of transgenic rice lines expressing the human lactoferrin gene
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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.
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.
참고문헌 (22)
- Arakawa T, "Improvements in human health through production of human milk proteins in transgenic food plants. in: Chemicals via Higher Plant Bioengineering", pp.1999, Kluwer Academic
- Bradford M, "A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding", Anal Biochem, 72, pp.248~254 (1976)
- Choi SM, "High expression of a human lactoferrin in transgenic tobacco cell cultures", Biotech Lett, 25, pp.213~218 (2003)
- Chong DKX, "Expression of full-length bioactive antimicrobial human lactoferrin in potato plants", Transgenic Res, 9, pp.71~78 (2000)
- Chong DKX, "Expression of the human milk protein β-casein in transgenic potato plants", Transgenic Res, 6, pp.289~296 (1997)
- Chu CC, "Establishment of an efficient medium for another culture of rice through comparative experiments on the nitrogen sources", Sci Sin, 18, pp.659~668 (1975)
- Hanson LA, "The role of breastfeeding in prevention of neonatal infection", Seminars in fetal & neonatal medicine, 7, pp.275~281 (2002)
- Lee TJ, "Partial resistance to bacterial wilt in transgenic tomato plants expressing antibacterial lactoferrin gene", J Amer Soc Hort Sci, 127, pp.158~164 (2002)
- Li Y, "Expression of a human lactoferrin N-lobe in Nicotiana benthamiana with potato virus X-based agroinfection", Biotechnol Lett, 26, pp.953~957 (2004)
- Liang Q, "Expression and characterization of human lactoferrin in yeast Saccharomyces cerevisiae", J Agric Food Chem, 41, pp.1800~1807 (1993)
- Lönnerdal B, "Recombinant human milk proteins-an opportunity and a challenge", Am J Clin Nutr, 63, pp.622S~626S (1996)
- Lönnerdal B, "Expression of human milk proteins in plants", J Am Clin Nutr, 21, pp.218S~221S (2002)
- Min SR, "Production of human lactoferrin in transgenic cell suspension cultures of sweet potato", Biologia Plantarum, 50, pp.131~134 (2006)
- Murray M, "Rapid isolation of molecular weight plant DNA", Nucleic Acid Res., 8, pp.4321~4325 (1980)
- Nandi S, "Expression of human lactoferrin in transgenic rice grains for the application in infant formula", Plant Sci, 163, pp.713~722 (2002)
- Nandi S, "Process development and economic evaluation of recombinant human lactoferrin expressed in rice grain", Transgenic Res, 14, pp.237~249 (2005)
- Platenburg GJ, "Expression of human lactoferrin in milk of transgenic mice", Transgenic Res, 3, pp.99~108 (1994)
- Philip MG, "Molecular plant biology volume one:a practical approach", pp.17~23 (2002)
- Salmon V, "Production of human lactoferrin in transgenic tobacco plants", Protein Express Purif, 13, pp.127~135 (1998)
- Simon R, "High frequency mobilization of gram negative bacterial replicons by the in vitro constructed Tn5-Mob transposon", Mol Gen Genet, 196, pp.413~420 (1984)
- Van Berkel PHC, "Large scale production of recombinant human lactoferrin in the milk of transgenic cows", Nature Biotechnol, 20, pp.484~487 (2002)
- Ward PP, "An inducible expression system for the production of human lactoferrin in Aspergillus nidulans", Gene, 122, pp.219~223 (1992)