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裂褶菌吲哚 - 3 - 甘油磷酸合成酶和磷酸核糖邻氨基苯甲酸异构酶编码DNA序列的分离。

Isolation of the DNA sequence coding indole-3-glycerol phosphate synthetase and phosphoribosylanthranilate isomerase of Schizophyllum commune.

作者信息

Muñoz-Rivas A M, Specht C A, Ullrich R C, Novotny C P

机构信息

Department of Botany, University of Vermont, Burlington 05405.

出版信息

Curr Genet. 1986;10(12):909-13. doi: 10.1007/BF00398288.

Abstract

A Schizophyllum gene library was made in plasmid pRK9. Plasmids from this library were tested for their ability to complement several auxotrophic mutations of Escherichia coli. The goal was to isolate a Schizophyllum auxotrophic gene that could be used to transform a corresponding Schizophyllum auxotrophic mutant to prototrophy. Complementation was observed only for E. coli trpC indole 3-glycerol phosphate synthetase (IGPS) and phosphoribosyl-anthranilate isomerase (PRAI) mutations. Plasmids with a Schizophyllum sequence coding for both IGPS and PRAI activities were recovered from E. coli transformants. Expression of the Schizophyllum gene (TRP1) in E. coli is probably dependent on the Serratia marcescens promoter of plasmid pRK9. The DNA sequence containing the Schizophyllum TRP1 gene was not obviously rearranged in cloning.

摘要

构建了裂褶菌基因文库,该文库以质粒pRK9为载体。对该文库中的质粒进行了测试,以检测其互补大肠杆菌几种营养缺陷型突变的能力。目的是分离出一个裂褶菌营养缺陷型基因,该基因可用于将相应的裂褶菌营养缺陷型突变体转化为原养型。仅在大肠杆菌色氨酸C吲哚-3-甘油磷酸合成酶(IGPS)和磷酸核糖基邻氨基苯甲酸异构酶(PRAI)突变中观察到互补作用。从大肠杆菌转化子中回收了编码IGPS和PRAI活性的裂褶菌序列的质粒。裂褶菌基因(TRP1)在大肠杆菌中的表达可能依赖于质粒pRK9的粘质沙雷氏菌启动子。含有裂褶菌TRP1基因的DNA序列在克隆过程中未明显重排。

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