生物活性人白细胞介素-18在转基因烟草植株中的表达与生产。
Expression and production of bioactive human interleukin-18 in transgenic tobacco plants.
作者信息
Zhang Bin, Yang Ying-Hua, Lin Yong-Ming, Rao Qing, Zheng Guo-Guang, Wu Ke-Fu
机构信息
National Laboratory of Experimental Hematology, Institute of Hematology & Blood Disease Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, PR China.
出版信息
Biotechnol Lett. 2003 Oct;25(19):1629-35. doi: 10.1023/a:1025638623770.
The cDNA of human interleukin-18 (hIL-18) was successfully inserted into the genome of tobacco plant, Nicotiana tabacum cv. NC-89, using Agrobacterium tumefaciens-mediated transformation. Insertion and translation of hIL-18 in transformants were confirmed by PCR, ELISA, and Western blot, respectively. The transformed extracts contained the recombinant hIL-18 protein up to 0.05% of total soluble protein. Activity of the recombinant hIL-18 in plant cells was confirmed by the induction of IFN-gamma on IL-18-responsive J6-1 cells by the extracts obtained from the transformants. The expression level of hIL-18 (351 ng g(-1) tobacco tissue) obtained in the present study may be sufficient to induce responses/effects in vivo.
利用根癌农杆菌介导的转化方法，人白细胞介素-18（hIL-18）的互补DNA（cDNA）成功插入烟草品种NC-89的基因组中。分别通过聚合酶链反应（PCR）、酶联免疫吸附测定（ELISA）和蛋白质免疫印迹法（Western blot）证实了转化体中hIL-18的插入和翻译。转化提取物中重组hIL-18蛋白的含量高达总可溶性蛋白的0.05%。通过转化体提取物对IL-18反应性J6-1细胞诱导γ-干扰素（IFN-γ），证实了植物细胞中重组hIL-18的活性。本研究中获得的hIL-18表达水平（351 ng g(-1)烟草组织）可能足以在体内诱导反应/效应。
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