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

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。...

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Published in广东农业科学 Vol. 43; no. 5; pp. 96 - 100
Main Author 任小平 蒲小明 沈会芳 张景欣 林壁润
Format Journal Article
LanguageChinese
Published 广东省农业有害生物预警防控中心,广东广州,510500%广东省农科院植物保护研究所,广东广州,510640 2016
Subjects
克隆
活性分析
表达
香蕉细菌性软腐病菌
克隆
香蕉细菌性软腐病菌
活性分析
表达
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ISSN1004-874X

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Summary: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。
Bibliography:44-1267/S
REN Xiao-ping, PU Xiao-ming, SHEN Hui-fang, ZHANG Jing-xin, LIN Bi-run （ 1, Forecast and Control Center for Agricultural Pest of Guangdong Provincie, Guangzhou 510500, China; 2.Plant Protection Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China ）
banana soft rot pathogen; gene cloning; expression; activity analysis
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,si
ISSN:1004-874X