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首页> 外文期刊>African Journal of Biotechnology >Expression and functional analysis of polygalacturonase gene Pcipg5 from the plant pathogen Phytophthora capsici
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Expression and functional analysis of polygalacturonase gene Pcipg5 from the plant pathogen Phytophthora capsici

机译：植物致病疫霉中聚半乳糖醛酸酶基因Pcipg5的表达及功能分析

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摘要

As one of the cell wall-degrading enzymes,?polygalacturonase involves in pathogenicity and virulence in a number of host pathogen interactions. In thepolygalacturonase multigene family of?Phytophthora capsici,?Pcipg5?was screened from the genomic library. The?Pcipg5 gene contains three putative active sites (179Asp, 200Asp, 201Asp) and one potential?N-glycosylation site (252Asn). PCIPG5, four active-site mutagenesis proteins and?N-glycosylation mutation were expressed inPichia pastoris?and purified by Ni-NTA Purification System.?Pcipg5 functions were determined by Western blot, RT-PCR and Northern blot. Transmission electron microscopy discovery individually in treated pepper leaves was used to determine the virulence of PCIPG5. The results show that?Pcipg5 can?encode a pathogenesis-related protein during?P. capsici?infection of pepper leaves and?degrade cell walls to produce necrotic lesions in treated pepper leaves. Meanwhile, the?N-glycosylation mutagenesis protein decreased the activity compared with PCIPG5. It indicates that the existence?N-glycosylation site in PCIPG5 plays a partial role in the activity of this enzyme.
机译：作为细胞壁降解酶之一，聚半乳糖醛酸酶在许多宿主病原体相互作用中涉及致病性和致病性。在辣椒疫霉菌的多半乳糖醛酸酶多基因家族中，从基因组文库中筛选了Pcipg5。 Pcipg5基因含有三个推定的活性位点（179Asp，200Asp，201Asp）和一个潜在的βN-糖基化位点（252Asn）。在巴斯德毕赤酵母中表达PCIPG5、4个活性位点诱变蛋白和N-糖基化突变，并通过Ni-NTA纯化系统纯化。通过Western印迹，RT-PCR和Northern印迹确定pcipg5的功能。分别在处理过的辣椒叶片中使用透射电子显微镜发现来确定PCIPG5的毒性。结果表明，Pcipg5可以在P期间编码与病程相关的蛋白质。辣椒叶片的辣椒素感染和降解的细胞壁在处理过的辣椒叶片中产生坏死性病变。同时，与PCIPG5相比，ΔN-糖基化诱变蛋白降低了活性。这表明PCIPG5中存在的βN-糖基化位点对该酶的活性起部分作用。

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