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构巢曲霉中的基因克隆:异柠檬酸裂解酶基因(acuD)的分离

Gene cloning in Aspergillus nidulans: isolation of the isocitrate lyase gene (acuD).

作者信息

Ballance D J, Turner G

出版信息

Mol Gen Genet. 1986 Feb;202(2):271-5. doi: 10.1007/BF00331649.

Abstract

An Aspergillus nidulans gene library was constructed in a high-frequency transformation vector, pDJB3, based on the Neurospora crassa pyr4 gene. This gene library was used to isolate the structural gene for isocitrate lyase (acuD) by complementation of a deficiency mutation following transformation of A. nidulans. Plasmids rescued in Escherichia coli were able to transform five different A. nidulans acuD mutants. Transformation using plasmids containing the cloned fragment resulted in integration at the acuD locus in six of nine transformants.

摘要

基于粗糙脉孢菌pyr4基因,在高频转化载体pDJB3中构建了构巢曲霉基因文库。该基因文库用于通过转化构巢曲霉后对缺陷突变的互补作用来分离异柠檬酸裂解酶(acuD)的结构基因。在大肠杆菌中拯救的质粒能够转化五种不同的构巢曲霉acuD突变体。使用含有克隆片段的质粒进行转化,在九个转化体中有六个在acuD位点发生整合。

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