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来自一种植物病原丝状真菌的促分裂原活化蛋白激酶编码cDNA的克隆与表达

Cloning and expression of cDNA encoding a mitogen-activated protein kinase from a phytopathogenic filamentous fungus.

作者信息

Li D, Rogers L, Kolattukudy P E

机构信息

Neurobiotechnology, The Ohio State University, Columbus, 43210, USA.

出版信息

Gene. 1997 Aug 22;195(2):161-6. doi: 10.1016/s0378-1119(97)00124-8.

Abstract

We have cloned a mitogen-activated protein kinase (MAPK) designated Fusarium solani f. sp. pisi mitogen-activated protein kinase (FsMAPK) from the phytopathogenic filamentous fungus F. solani f. sp. pisi T8 strain. A single open reading frame (ORF) of 1068 bp encoding a polypeptide of 355 amino acids (aa) with a predicted molecular mass of 41,194 Da was found in the cloned 1583-bp cDNA insert. FsMAPK is highly homologous to SPK1 of fission yeast, FUS3 of budding yeast, MsERK1 of alfalfa, Sur-1 of nematode, and hERK1 of human. That this gene is expressed in F. solani f. sp. pisi was shown by the finding that immunoblot of the fungal extracts with anti-FsMAPK antibodies (Ab) raised in a rabbit against the FsMAPK, expressed in Escherichia coli (E. coli), detected the corresponding protein. DNA blot analysis indicated that Fsmapk is present as a single copy in the fungal genome.

摘要

我们从植物病原丝状真菌豌豆尖镰孢菌(Fusarium solani f. sp. pisi)T8菌株中克隆了一种丝裂原活化蛋白激酶(MAPK),命名为豌豆尖镰孢菌丝裂原活化蛋白激酶(FsMAPK)。在克隆的1583 bp cDNA插入片段中发现了一个1068 bp的单一开放阅读框(ORF),其编码一个由355个氨基酸(aa)组成的多肽,预测分子量为41,194 Da。FsMAPK与裂殖酵母的SPK1、芽殖酵母的FUS3、苜蓿的MsERK1、线虫的Sur-1以及人类的hERK1高度同源。用针对在大肠杆菌(E. coli)中表达的FsMAPK在兔体内产生的抗FsMAPK抗体(Ab)对真菌提取物进行免疫印迹分析,检测到了相应的蛋白质,这表明该基因在豌豆尖镰孢菌中表达。DNA印迹分析表明,Fsmapk在真菌基因组中以单拷贝形式存在。

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