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水稻白叶枯病菌harpin基因的克隆与表达
CLONING AND EXPRESSING A harpin GENE FROM Xanthomonas oryzae pv.oryzae
【摘要】 利用 PCR方法从水稻黄单胞菌白叶枯致病变种 (Xanthomonas oryzae pv.oryzae,Xoo) Jxo III菌株中克隆到编码诱导植物过敏性反应激发子的基因 hrf A。同源性比较发现其推测产物与水稻白叶枯病菌菲律宾小种 Pxo86的 Hpa1有 97.2 %的序列一致性 ,比 Hpa1少了一个 - GGG- GG-氨基酸残基序列重复。基因经 Nde I和 Bam HI双酶切后连到含 T7启动子的高表达载体 p ET30 a(+ )上构建重组质粒 p HRF4 ,并转化宿主菌 BL2 1(DE3)产生表达菌株 BL HR4。经过 IPTG诱导之后 ,BL HR4产生一分子量为 15 .6k D的蛋白质。研究表明 ,该蛋白在性质与功能上类似于其它已发现的 harpins,即能够在烟草上引起典型的过敏性反应 ,富含甘氨酸、热稳定以及对蛋白酶敏感。因此 ,我们把该蛋白定名为 harpin Xoo。harpin Xoo还具有诱导植物抗病性的功能。
【Abstract】 A hrfA gene which encode plant hypersensitive response elicitor was cloned from Xanthomonas oryzae pv. oryzae isolate JxoIII by PCR. It encodes a predicted 140 amino acids protein that is glycine rich. Homology research showed that the protein is 97.2% identical with the Hpa1 of Xanthomonas oryzae pv. oryzae (Pxo86), but the number of the GGG GG repeats is one less. In Escherichia coli , BLHR4 strain overexpressing hrfA , the 15.6 kD protein fractionated from E.coli has been identified as the harpin Xoo . Like other harpins, harpin Xoo eli cits a typical HR in tobacco leaves. It is heat stable and appear highly sensitive to protease. Moreover, harpin Xoo has the function of inducing plant disease resistance.
【Key words】 Xanthomonas oryzae pv. oryzae; hypersensitive response; hrfA; harpin Xoo; induced disease resistance;
- 【文献出处】 植物病理学报 ,Acta Phytopathologica Sinica , 编辑部邮箱 ,2001年04期
- 【分类号】S435.111;S432.4
- 【被引频次】132
- 【下载频次】575