Kanvinde L, Anding H, Ozin L, Miller I S, Sastry G R
Mol Gen Genet. 1987 Mar;206(3):460-4. doi: 10.1007/BF00428886.
Plasmids containing hybrid genes, in which different Klebsiella pneumoniae nif (nitrogen-fixation) promoters were fused with the structural part of the Escherichia coli lac operon, were introduced into a double auxotrophic derivative of Agrobacterium tumefaciens C58. A study of their expression in the new host was made simple by the inherent inability of A. tumefaciens C58 to produce beta-galactosidase unless provided with the wild-type lac operon of E. coli. As shown by quantitative measurements of the enzyme, all K. pneumoniae promoters were expressed well in A. tumefaciens C58, even under conditions known to repress them. It also has been shown that the activity of K. pneumoniae nif A is essential for the expression of nifHDK even when introduced into A. tumefaciens. After entering the new host the plasmids, the nif genes and the fusion alleles contained in them, remained stable. Possible mechanisms responsible for the constitutive behaviour of nif promoters in A. tumefaciens are discussed.
含有杂交基因的质粒被导入根癌土壤杆菌C58的双营养缺陷型衍生物中,这些杂交基因是将不同的肺炎克雷伯氏菌固氮(nif)启动子与大肠杆菌乳糖操纵子的结构部分融合而成。由于根癌土壤杆菌C58自身无法产生β-半乳糖苷酶,除非提供大肠杆菌的野生型乳糖操纵子,这使得对它们在新宿主中的表达研究变得简单。通过对该酶的定量测量表明,即使在已知会抑制它们的条件下,所有肺炎克雷伯氏菌启动子在根癌土壤杆菌C58中都能良好表达。研究还表明,即使将肺炎克雷伯氏菌nif A导入根癌土壤杆菌,其活性对于nifHDK的表达也是必不可少的。质粒进入新宿主后,其中所含的nif基因和融合等位基因保持稳定。文中讨论了根癌土壤杆菌中nif启动子组成型行为的可能机制。