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首页> 外文期刊>Journal of natural toxins >Cloning, expression, and biological activity of recombinant alpha-cinnamomin: toxicity to cranberry and other plant species.
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Cloning, expression, and biological activity of recombinant alpha-cinnamomin: toxicity to cranberry and other plant species.

机译：重组α-肉桂酸酯的克隆，表达和生物学活性：对蔓越莓和其他植物物种的毒性。

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摘要

Elicitins produced by the pathogenic fungi Phytophthora are known to exhibit The elicitin cinnamomin is of nonspecific toxicity to different solanaceous plant species. particular interest for its potential role in the hypersensitive-like cell death and in the biological response of cranberry plants to the fungal pathogen Phytophthora cinnamomi. In order to understand the biochemical steps of the Phytophthora root rot disease in cranberry, we investigated the alpha-cinnamomin-induced plant responses. Toxicity of alpha-cinnamomin, which shows a high degree of sequence homology to the alpha-elicitin group, was tested on Vaccinum macrocarpon, Nicotiana tabacum, Capsicum annuum, Lycopersicon esculentum, Lactuca sativa, and Phaseolus vulgaris plants. Gene corresponding to alpha-cinnamomin gene fused with maltose binding protein gene, was cloned into a pMALTEV expression vector, which was transformed into E. coli cells. Cells containing alpha-cinnamomin clones were cultured and extracted protein was purified on a maltose binding protein affinity column. Biological activity of alpha-cinnamomin fusion protein was examined on propagated plants and cuttings. In cranberry plants treated with cinnamomin necrotic hypersensitive-like response in the proximal areas of the leaf lamina of lower plant leaves was observed after 48-72 hr of incubation. Limited leaf necrosis observed days after application of low amounts of recombinant cinnamomin directly on the leaves of other plants indicates that the recombinant protein might be functioning as a toxin, capable of inducing aging accompanied by plant cell death.
机译：已知由致病性真菌疫霉属产生的抑肽素显示出elicitin cinnamomin对不同茄科植物具有非特异性毒性。它在类似过敏的细胞死亡以及蔓越莓植物对真菌病原菌Phytophthora cinnamomi的生物反应中的潜在作用特别令人感兴趣。为了了解蔓越莓疫霉根腐病的生化步骤，我们研究了α-肉桂胺诱导的植物反应。在大花牛痘苗，烟草，辣椒，番茄，番茄，菜豆和菜豆上测试了α-肉桂胺的毒性，该毒素与α-诱导素组具有高度的序列同源性。将与麦芽糖结合蛋白基因融合的对应于α-肉桂酰胺基因的基因克隆到pMALTEV表达载体中，将其转化到大肠杆菌细胞中。培养含有α-肉桂酰胺克隆的细胞，并在麦芽糖结合蛋白亲和柱上纯化提取的蛋白。在繁殖的植物和插条上检查了α-肉桂酰胺融合蛋白的生物活性。孵育48-72小时后，在下部植物叶片的叶片叶片近端区域中，用肉桂酸坏死性高敏样反应处理的蔓越莓植物被观察到。直接在其他植物的叶子上直接施用少量的重组肉桂黄素后数天观察到有限的叶子坏死，表明重组蛋白可能起毒素的作用，能够诱导衰老并伴随植物细胞死亡。

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