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首页> 外文期刊>Breeding science >Production and characterization of recombinant human lactoferrin in transgenic Javanica rice.
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Production and characterization of recombinant human lactoferrin in transgenic Javanica rice.

机译：转基因爪哇稻中重组人乳铁蛋白的生产和表征。

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摘要

The objective of the present study was to develop a functional crop that could contribute to the maintenance and improvement of human health by the introduction of the human lactoferrin (hLF) gene. Lactoferrin is an 80-kDa iron-binding glycoprotein that has been considered to play many biological roles, including the regulation of iron absorption, protection against microbial and virus infection, stimulation of the immune system and cellular growth promotion. We introduced two different constructs containing either the native signal peptide from human lactoferrin (pIG211) or the signal peptide from rice glutelin (pIG200) fused to mature human lactoferrin into Javanica rice cv. Rojolele by using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the maize ubiquitin-1 promoter was detected in all the tissues of the transgenic plants. We found that the transgenic rice plants IG200R produced a considerable amount of recombinant hLF (rhLF) in seeds, accounting for approximately 15% of the total soluble protein (TSP). RhLF was purified from mature seeds by cation-exchange chromatography. Amino acid sequencing confirmed that the N-terminal sequences of rhLF for both constructs were identical with those of native LF from human milk. Rice rhLF showed a slightly smaller molecular weight than the native hLF. Immunofluorescence microscopy revealed that rhLF was located in the intracellular and intercellular regions of endosperm cells. Protein extracts from transgenic rice seeds exhibited an antibacterial activity against Bacillus subtilis ATCC6633. The use of signal peptides and a constitutive ubiquitin-1 promoter for successful production of transgenic Javanica rice expressing a high level of rhLF was examined..
机译：本研究的目的是开发一种功能性作物，通过引入人乳铁蛋白（hLF）基因，可以有助于维持和改善人类健康。乳铁蛋白是一种80 kDa的铁结合糖蛋白，被认为具有许多生物学作用，包括调节铁的吸收，防止微生物和病毒感染，刺激免疫系统和促进细胞生长。我们引入了两种不同的构建体，它们包含来自人乳铁蛋白的天然信号肽（pIG211）或来自将成熟人乳铁蛋白融合到爪哇稻米中的稻谷蛋白（pIG200）的信号肽。 Rojolele通过使用农杆菌介导的转化系统。在转基因植物的所有组织中检测到在玉米泛素-1启动子控制下的hLF基因的表达。我们发现转基因水稻植物IG200R在种子中产生了大量重组hLF（rhLF），约占总可溶性蛋白（TSP）的15％。通过阳离子交换色谱法从成熟种子中纯化RhLF。氨基酸测序证实，两种构建体的rhLF的N末端序列与人乳的天然LF的N末端序列相同。水稻rhLF的分子量略小于天然hLF。免疫荧光显微镜检查显示，rhLF位于胚乳细胞的细胞内和细胞间区域。来自转基因水稻种子的蛋白质提取物表现出对枯草芽孢杆菌ATCC6633的抗菌活性。检验了信号肽和组成型泛素-1启动子在成功生产表达高水平rhLF的转基因爪哇稻中的用途。

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