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拟南芥中一种类PR5蛋白的克隆与表达:细菌表达的该蛋白对真菌生长的抑制作用

Cloning and expression of a PR5-like protein from Arabidopsis: inhibition of fungal growth by bacterially expressed protein.

作者信息

Hu X, Reddy A S

机构信息

Department of Biology, Colorado State University, Fort Collins 80523, USA.

出版信息

Plant Mol Biol. 1997 Aug;34(6):949-59. doi: 10.1023/a:1005893119263.

Abstract

Pathogenesis-related (PR)-5 proteins are a family of proteins that are induced by different phytopathogens in many plants and share significant sequence similarity with thaumatin. We isolated a complementary DNA (ATLP-3) encoding a PR5-like protein from Arabidopsis which is distinct from two other previously reported PR5 cDNAs from the same plant species. The predicted ATLP-3 protein with its amino-terminal signal sequence is 245 amino acids in length and is acidic with a pl of 4.8. The deduced amino acid sequence of ATLP-3 shows significant sequence similarity with PR5 and thaumatin-like proteins from Arabidopsis and other plants and contains a putative signal sequence at the amino-terminus. The expression of ATLP-3 and a related gene (ATLP-1) that we previously isolated from Arabidopsis was induced by pathogen infection and salicylic acid, a known inducer of pathogenesis-related genes. Southern blot analysis indicates that the ATLP-1 and ATLP-3 are coded by single-copy genes. To study the effect of ATLP-1 and ATLP-3 proteins on fungal growth, the cDNA regions corresponding to putative mature protein were expressed in Escherichia coli and the cDNA encoded proteins were purified. ATLP-1 and ATLP-3 proteins cross-reacted with anti-osmotin and anti-zeamatin antibodies. ATLP-3 protein showed antifungal activity against several fungal pathogens suggesting that ATLP-3 may be involved in plant defense against fungal pathogens.

摘要

病程相关(PR)-5蛋白是一类在许多植物中由不同植物病原体诱导产生的蛋白质,与奇异果甜蛋白具有显著的序列相似性。我们从拟南芥中分离出一个编码PR5样蛋白的互补DNA(ATLP-3),它与同一植物物种中先前报道的另外两个PR5 cDNA不同。预测的ATLP-3蛋白连同其氨基末端信号序列长度为245个氨基酸,呈酸性,等电点为4.8。推导的ATLP-3氨基酸序列与来自拟南芥和其他植物的PR5以及类奇异果甜蛋白具有显著的序列相似性,并且在氨基末端含有一个推定的信号序列。我们先前从拟南芥中分离出的ATLP-3和一个相关基因(ATLP-1)的表达受病原体感染和水杨酸(一种已知的病程相关基因诱导剂)诱导。Southern印迹分析表明,ATLP-1和ATLP-3由单拷贝基因编码。为了研究ATLP-1和ATLP-3蛋白对真菌生长的影响,将对应于推定成熟蛋白的cDNA区域在大肠杆菌中表达,并纯化cDNA编码的蛋白。ATLP-1和ATLP-3蛋白与抗渗透素和抗玉米素抗体发生交叉反应。ATLP-3蛋白对几种真菌病原体表现出抗真菌活性,表明ATLP-3可能参与植物对真菌病原体的防御。

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