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编码NADP连接型谷氨酸脱氢酶的构巢曲霉gdhA基因的分离与鉴定。

Isolation and identification of the Aspergillus nidulans gdhA gene encoding NADP-linked glutamate dehydrogenase.

作者信息

Gurr S J, Hawkins A R, Drainas C, Kinghorn J R

机构信息

Department of Biochemistry and Microbiology, University of St. Andrews, St. Andrews Fife, UK.

出版信息

Curr Genet. 1986;10(10):761-6. doi: 10.1007/BF00405099.

Abstract

The Neurospora crassa am gene was used as a heterologous probe to identify clones from two independently constructed Aspergillus nidulans gene libraries. These clones have a common HindIII 1.85 kb fragment. This A. nidulans nucleotide stretch hybridises to a N. crassa 2.7 kb BamHI fragment of wild type DNA but not to a co-migrating fragment from the DNA of the N. crassa am132 deletion mutant. One A. nidulans clone was shown to complement the N. crasse am132 deletion strain. The N. crassa transformants show low levels (approximately 5%) of heterologous glutamate dehydrogenase activity. The A. nidulans gdhA gene was found to locate in N. crassa at both the "homologous" (i.e. am) site as well as non-homologous sites. Partial nucleotide analysis of the fragment has revealed the 5' end of the locus and considerable homology with the N. crassa am gene. We concluded that we have cloned the A. nidulans gdhA gene.

摘要

粗糙脉孢菌的am基因被用作异源探针,以从两个独立构建的构巢曲霉基因文库中鉴定克隆。这些克隆有一个共同的HindIII 1.85 kb片段。这个构巢曲霉的核苷酸片段与野生型粗糙脉孢菌DNA的一个2.7 kb BamHI片段杂交,但不与粗糙脉孢菌am132缺失突变体DNA的共迁移片段杂交。一个构巢曲霉克隆被证明可以互补粗糙脉孢菌am132缺失菌株。粗糙脉孢菌转化体显示出低水平(约5%)的异源谷氨酸脱氢酶活性。发现构巢曲霉的gdhA基因定位于粗糙脉孢菌的“同源”(即am)位点以及非同源位点。对该片段的部分核苷酸分析揭示了该基因座的5'端,并与粗糙脉孢菌的am基因有相当的同源性。我们得出结论,我们已经克隆了构巢曲霉的gdhA基因。

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