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子囊菌大孢粪壳菌的ura5基因:分子克隆、特性鉴定及在大肠杆菌中的表达

The ura5 gene of the ascomycete Sordaria macrospora: molecular cloning, characterization and expression in Escherichia coli.

作者信息

Le Chevanton L, Leblon G

机构信息

Laboratoire Interactions Moléculaires Génomiques, Université Paris-Sud, Orsay, France.

出版信息

Gene. 1989 Apr 15;77(1):39-49. doi: 10.1016/0378-1119(89)90357-0.

Abstract

We cloned the ura5 gene coding for the orotate phosphoribosyl transferase from the ascomycete Sordaria macrospora by heterologous probing of a Sordaria genomic DNA library with the corresponding Podospora anserina sequence. The Sordaria gene was expressed in an Escherichia coli pyrE mutant strain defective for the same enzyme, and expression was shown to be promoted by plasmid sequences. The nucleotide sequence of the 1246-bp DNA fragment encompassing the region of homology with the Podospora gene has been determined. This sequence contains an open reading frame of 699 nucleotides. The deduced amino acid sequence shows 72% similarity with the corresponding Podospora protein.

摘要

我们通过用相应的嗜鸟粪层孔菌序列对大孢粪壳菌基因组DNA文库进行异源杂交,克隆了编码乳清酸磷酸核糖转移酶的ura5基因。该粪壳菌基因在一株对同一种酶有缺陷的大肠杆菌pyrE突变菌株中表达,并且表达受到质粒序列的促进。已经确定了包含与嗜鸟粪层孔菌基因同源区域的1246 bp DNA片段的核苷酸序列。该序列包含一个699个核苷酸的开放阅读框。推导的氨基酸序列与相应的嗜鸟粪层孔菌蛋白显示出72%的相似性。

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