Gayda D J, Leathers T D, Noti J D, Smith F J, Smith J M, Subrahmanyam C S, Umbarger H E
J Bacteriol. 1980 May;142(2):556-67. doi: 10.1128/jb.142.2.556-567.1980.
A strain of Escherichia coli K-12 containing a deletion extending from early in the ilvE gene toward the ilvG gene was shown to exhibit a higher expression of the downstream genes, ilvD and ilvA, than did an ilv+ strain. The elevated expression was under apparently normal ilv-specific control, however. The deletion was transferred to the ilv region of lamba h80dilv and shown by restriction endonuclease and heteroduplex analysis to extend through the deoxyribonucleic acid (DNA) shown, in the preceding paper (C. S. Subrahmanyam, G. M. McCorkle, and H. E. Umbarget, J. Bacteriol 142:547--555, 1980), to contain the ilvO determinant. The deletion was also transferred to an ilv-lac fusion strain and shown to cause an increase in beta-galactosidase formation while allowing retention of ilv-specific control. Transducing phages excised from these fusion strains with and without the ilvO determinant were compared. The phage carrying the ilvO+ determinant contained ilv DNA extending only into but not through the ilvG gene. It did not exhibit an ilv-specific control of beta-galactosidase formation. The phage carrying the deletion of ilvO but containing ilv DNA extending beyond the ilvG gene exhibited ilv-specific control of beta-galactosidase formation. It was concluded that the multivalently controlled ilv-specific promoter affecting ilv operon expression lies upstream from ilvG and that the ilvO region in the wild-type K-12 strain is a region of polarity preventing ilvG expression and reducing ilvEDA expression.
一株含有从ilvE基因早期延伸至ilvG基因的缺失的大肠杆菌K-12菌株,与ilv⁺菌株相比,显示出下游基因ilvD和ilvA的表达更高。然而,这种升高的表达处于明显正常的ilv特异性控制之下。该缺失被转移到λh80dilv的ilv区域,并通过限制性内切酶和异源双链分析表明,它延伸穿过在前一篇论文(C.S.Subrahmanyam、G.M.McCorkle和H.E.Umbarget,《细菌学杂志》142:547 - 555,1980)中所示的包含ilvO决定簇的脱氧核糖核酸(DNA)。该缺失也被转移到一个ilv-lac融合菌株中,并显示会导致β-半乳糖苷酶形成增加,同时允许保留ilv特异性控制。比较了从这些有和没有ilvO决定簇的融合菌株中切除的转导噬菌体。携带ilvO⁺决定簇的噬菌体所含的ilv DNA仅延伸到ilvG基因内但未穿过该基因。它没有表现出对β-半乳糖苷酶形成的ilv特异性控制。携带ilvO缺失但含有延伸超过ilvG基因的ilv DNA的噬菌体表现出对β-半乳糖苷酶形成的ilv特异性控制。得出的结论是,影响ilv操纵子表达的多价控制的ilv特异性启动子位于ilvG上游,并且野生型K-12菌株中的ilvO区域是一个极性区域,可阻止ilvG表达并降低ilvEDA表达。