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香蕉细菌性软腐病菌细胞壁降解酶pelD、pelE基因的克隆表达及活性分析

Cloning and expression analysis of cell wall degrading enzyme genes from banana soft rot pathogen

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【作者】 任小平蒲小明沈会芳张景欣林壁润

【Author】 REN Xiao-ping;PU Xiao-ming;SHEN Hui-fang;ZHANG Jing-xin;LIN Bi-run;Forecast and Control Center for Agricultural Pest of Guangdong Provincie;Plant Protection Institute,Guangdong Academy of Agricultural Sciences;

【机构】 广东省农业有害生物预警防控中心广东省农科院植物保护研究所

【摘要】 细胞壁降解酶与植物病害症状有重要的相关性。根据已知软腐病细菌细胞壁降解酶基因序列,设计两对特异性引物,对香蕉细菌性软腐病菌进行PCR克隆,获得具有完整阅读框的基因pelD、pelE序列。通过构建重组表达载体pET28b(+)-pelD和pET28b(+)-pelE,重组表达载体转化子在IPTG诱导下,经SDSPAG分析,获得了pelD和pelE的表达蛋白。将两种表达蛋白接种于7~8片叶的粉蕉假茎和离体叶片上,发现接种pelE表达蛋白的粉蕉假茎和叶片出现了腐烂症状,但接种pelD表达蛋白没有引起发病症状。

【Abstract】 Cell wall degrading enzymes had important relationship with the symptoms of plant diseases. Two pairs of specific primers were designed for amplification and cloning gene sequences of Banana soft rot pathogen according to reported cell wall degrading enzyme genes. A complete reading frame sequence of gene pelD and pelE were obtained by cloning technology. The expression vector pET28b (+) was used to investigate the pelD and pelE gene expression.According to the results of SDS-PAGE,under the inducing of IPTG,transformants of recombinant expression vectors expressed much protein. After the purified protein was inoculated into banana false stem and leaves in vitro,similar rot symptoms were presented compared to Banana bacterial soft rot pathogen,the purified protein from vector pET28b (+) -pelE after expressing 12 h appeared serious rot,while the purified protein from vector pET28b (+) –pelD did not cause rot in plant.

【基金】 国家自然科学基金(31300118);广东省国际合作项目(2012B050500014);广东省自然科学基金(S2013010012424;2015A030312002)
  • 【文献出处】 广东农业科学 ,Guangdong Agricultural Sciences , 编辑部邮箱 ,2016年05期
  • 【分类号】S436.68
  • 【被引频次】3
  • 【下载频次】117
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