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日本麒麟啤酒公司基础技术研究所和北海道大学农学部助教授山冈直人等小组在世界上首次从大豆中克隆出作为诱导植物抗病性诱发剂的β葡聚糖受体基因。麒麟啤酒公司打算在该公司农业事业的主要产品花卉上大量表达,开发对病原菌感染应答快的抗菌植物。这项成果于1996年3月27日在鹿儿岛召开的日本植物生理学会上发表。该公司从40kg大豆根中取出存在诱发剂受体的细胞膜部分,经过6个步骤进行纯化。纯化产物有与β葡聚糖结合的机能,β葡聚糖含有作为诱发剂起作用的β-1,3和β-1,6键,平均分子量约为1万。得到的蛋白分子量为70kD,以这种蛋白的部分序列为基础,进行PCR,得到cDNA。用烟草培养细胞BY2表达后取出膜部分,除确认有与β葡聚糖的结合机能外,还确认了利用小鼠多克隆抗体抑制基质结合和植物抗菌素在培养基中的累积。另外,用免疫电子显微镜观察受体存在于细胞膜表面。
Japan Kirin Brewery Institute of Basic Technology and Hokkaido University Faculty of Agriculture Assistant Professor Yamanaka Yamaguchi and other groups for the first time in the world cloned from soybeans as induced plant resistance to β-glucan receptor gene. Kirin Brewery intends to express a great deal of flowers on the company’s main agricultural product and develop antimicrobial plants that respond quickly to pathogen infections. The result was presented at the Japanese Society of Plant Physiology, held on March 27, 1996, in Kagoshima. The company removed the cell membrane portion of the elicitor receptor from 40 kg of soy root and purified it in six steps. The purified product has the function of binding to β-glucan which contains β-1,3 and β-1,6 bonds acting as inducers and has an average molecular weight of about 10,000. The resulting protein has a molecular weight of 70 kD. Based on the partial sequence of this protein, PCR was performed to obtain cDNA. It was also confirmed that the use of mouse polyclonal antibody inhibited the binding of the matrix and the accumulation of plant antibiotics in the medium, in addition to confirming the binding function to β-glucan, by taking out the membrane fraction after expressing BY2 cells in tobacco. In addition, the presence of the receptor on the cell membrane surface was observed with an immunoelectron microscope.