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鉴定镰孢菌属中新的半乳糖氧化酶基因

Identification of new galactose oxidase genes in Fusarium spp.

机构信息

Departament of Biochemistry, Universidade Estadual de Maringá, Maringá, PR, Brazil.

出版信息

J Basic Microbiol. 2010 Dec;50(6):527-37. doi: 10.1002/jobm.201000078.

Abstract

Galactose oxidase (GO) converts galactose to an aldehyde and has several biotechnological applications, including cancer diagnosis. It is mainly produced by Fusarium austroamericanum but is also produced by Fusarium acuminatum and by isolates of the Fusarium graminearum and Gibberella fujikuroi complexes. The F. austroamericanum GO gaoA gene has been cloned, but the GO genes from other secreting species have not been characterized. Problems associated with the F. austroamericanum GO such as high pI and low catalytic efficiency and thermostability, and the difficult purification process makes the search for homologous genes attractive. In this work, the GO genes from Fusarium verticillioides and Fusarium subglutinans, two species of the G. fujikuroi complex, were cloned, sequenced, and analyzed. New GO genes were found in databases and were used to construct a phylogenetic tree, which revealed the existence of three orthologous lineages of GO genes in Fusarium spp. In addition, RT-PCR analyses revealed that the new GO cloned gene may be endogenously expressed in F. subglutinans but not in F. verticillioides, in the used culture conditions.

摘要

半乳糖氧化酶(GO)将半乳糖转化为醛,具有多种生物技术应用,包括癌症诊断。它主要由美洲镰孢菌产生,但也由尖孢镰孢菌和禾谷镰孢菌和藤仓镰刀菌复合体的分离株产生。已克隆出美洲镰孢菌 GO 的 gaoA 基因,但其他分泌物种的 GO 基因尚未得到表征。与美洲镰孢菌 GO 相关的问题,如高等电点、低催化效率和热稳定性,以及困难的纯化过程,使得寻找同源基因具有吸引力。在这项工作中,从藤仓镰刀菌和禾谷镰刀菌两个藤仓镰刀菌复合体的物种中克隆、测序和分析了 GO 基因。在数据库中发现了新的 GO 基因,并用于构建系统发育树,该树揭示了在镰刀菌属中存在三个 GO 基因的直系同源谱系。此外,RT-PCR 分析表明,在使用的培养条件下,新克隆的 GO 基因可能在 F. subglutinans 中内源性表达,但在 F. verticillioides 中不表达。

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