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通过转化导入粗糙脉孢菌后构巢曲霉QUTE基因的诱导表达。

Induced expression of the Aspergillus nidulans QUTE gene introduced by transformation into Neurospora crassa.

作者信息

Hiett K L, Case M E

机构信息

Department of Genetics, University of Georgia, Athens 30602.

出版信息

Mol Gen Genet. 1990 Jul;222(2-3):201-5. doi: 10.1007/BF00633818.

Abstract

The qa-2 gene of Neurospora crassa encodes catabolic dehydroquinase which catabolizes dehydroquinic acid to dehydroshikimic acid. The QUTE gene of Aspergillus nidulans corresponds to the qa-2 gene of N. crassa. The plasmid pEH1 containing the QUTE gene from A. nidulans was used to transform a qa-2- strain of N. crassa. In Southern blot analyses, DNAs isolated from these transformants hybridized specifically to the QUTE gene probe. Northern blot analyses indicated that QUTE mRNA was produced in the transformants. The functional integrity of the QUTE gene in N. crassa was indicated by transformants which had regained the ability to grow on quinic acid as sole carbon source. Enzyme assays indicated that the specific activities of catabolic dehydroquinase induced by quinic acid in the transformants ranged from 4% to 32% of that induced in wild-type N. crassa. The evidence that the QUTE structural gene of A. nidulans is inducible when introduced into the N. crassa genome implies that the N. crassa qa activator protein can recognize, at least to a limited extent, DNA binding sequences 5' to the QUTE gene.

摘要

粗糙脉孢菌的qa - 2基因编码分解代谢脱氢奎尼酸酶,该酶将脱氢奎尼酸分解为脱氢莽草酸。构巢曲霉的QUTE基因与粗糙脉孢菌的qa - 2基因相对应。含有构巢曲霉QUTE基因的质粒pEH1被用于转化粗糙脉孢菌的qa - 2 -菌株。在Southern印迹分析中,从这些转化体中分离的DNA与QUTE基因探针特异性杂交。Northern印迹分析表明转化体中产生了QUTE mRNA。在以奎尼酸作为唯一碳源时能够恢复生长能力的转化体表明了粗糙脉孢菌中QUTE基因的功能完整性。酶活性测定表明,奎尼酸在转化体中诱导产生的分解代谢脱氢奎尼酸的比活性为野生型粗糙脉孢菌中诱导产生的比活性的4%至32%。构巢曲霉的QUTE结构基因导入粗糙脉孢菌基因组后具有可诱导性,这一证据表明粗糙脉孢菌的qa激活蛋白至少在一定程度上能够识别QUTE基因5'端的DNA结合序列。

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