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麦角菌甘油醛-3-磷酸脱氢酶基因:克隆、表征及其在优化显性选择系统中的应用

The Claviceps purpurea glyceraldehyde-3-phosphate dehydrogenase gene: cloning, characterization, and use for the improvement of a dominant selection system.

作者信息

Jungehülsing U, Arntz C, Smit R, Tudzynski P

机构信息

Allgemeine Botanik/Mikrobiologie, Westfälische Wilhelms-Universität, Münster, Germany.

出版信息

Curr Genet. 1994 Feb;25(2):101-6. doi: 10.1007/BF00309533.

Abstract

The GPD 1 gene of Claviceps purpurea coding for glyceraldehyde-3-phosphate dehydrogenase was cloned and sequenced, including 1,800 bp of its 5' upstream region. This gene shows an identical structure to the gpd gene of Podospora anserina and Cryphonectria parasitica (one intron at an identical position) with high homology at both the DNA and amino-acid levels. Two fragments of the promoter spanning from the ATG to -500 bp and to -1,400 bp were fused to the phleomycin-resistance gene. Both constructs transformed C. purpurea at a high rate. The enhanced expression of the long vector construct indicates the presence of additional elements between -500 bp and -1,400 bp upstream of the initiation codon.

摘要

对编码甘油醛-3-磷酸脱氢酶的麦角菌(Claviceps purpurea)的GPD 1基因进行了克隆和测序,包括其5'上游区域的1800 bp。该基因与栗疫霉(Podospora anserina)和寄生隐孢壳菌(Cryphonectria parasitica)的gpd基因结构相同(在相同位置有一个内含子),在DNA和氨基酸水平上具有高度同源性。将启动子的两个片段,即从ATG到-500 bp和到-1400 bp的片段,与博来霉素抗性基因融合。两种构建体都以高转化率转化了麦角菌。长载体构建体的增强表达表明在起始密码子上游-500 bp和-1400 bp之间存在其他元件。

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