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利用基因融合技术研究大肠杆菌K-12的负调控基因uidR的表达。

The use of gene fusions to study the expression of uidR, a negative regulatory gene of Escherichia coli K-12.

作者信息

Blanco C, Mata-Gilsinger M, Ritzenthaler P

出版信息

Gene. 1985;36(1-2):159-67. doi: 10.1016/0378-1119(85)90080-0.

Abstract

The uidR regulatory gene of Escherichia coli codes for a repressor molecule that negatively controls the expression of the uidA gene. The uidR gene was fused in front of the lacZ gene in vitro on plasmid cloning vectors developed by Casadaban et al. [J. Bacteriol. 143 (1980) 971-980. The transcriptional direction of uidR was deduced from the restriction pattern and the phenotypic properties of the uidR-lacZ fusion plasmids. The gene is transcribed counterclockwise on the standard E. coli map, as is the uidA gene. The uidR-lacZ fusions were also used to examine the regulation of expression from the uidR promoter. It was observed that the uidR gene expression is repressed by its own product and is sensitive to catabolite control. The uidR-lacZ-encoded proteins of various sizes were isolated but attempts to obtain hybrid molecules possessing, in a single polypeptide, both the beta-galactosidase and UidR repressor activities were unsuccessful.

摘要

大肠杆菌的uidR调控基因编码一种阻遏分子,该分子对uidA基因的表达进行负调控。在卡萨达班等人开发的质粒克隆载体上,体外将uidR基因融合到lacZ基因的前面[《细菌学杂志》143(1980)971 - 980]。根据uidR - lacZ融合质粒的限制酶切图谱和表型特性推断出uidR的转录方向。该基因在标准大肠杆菌图谱上逆时针转录,uidA基因也是如此。uidR - lacZ融合体也用于检测uidR启动子的表达调控。观察到uidR基因的表达受到其自身产物的抑制,并且对分解代谢物控制敏感。分离出了各种大小的uidR - lacZ编码蛋白,但试图获得在单个多肽中同时具有β - 半乳糖苷酶和UidR阻遏活性的杂交分子未成功。

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