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。
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.
出处
《广东农业科学》
CAS
2016年第5期96-100,F0003,共6页
Guangdong Agricultural Sciences
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)
关键词
香蕉细菌性软腐病菌
克隆
表达
活性分析
banana soft rot pathogen
gene cloning
expression
activity analysis
