作者: Daniela Jacob , Astrid Lewin , Beate Meister , Bernd Appel
关键词: Transcription (biology) 、 Bacterial transcription 、 Biology 、 Gene expression 、 Genetics 、 Response element 、 Cauliflower mosaic virus 、 Gene 、 Promoter 、 Expression vector
摘要: During evolution the promoter elements from prokaryotes and eukaryotes have developed differently with regard to their sequence structure, implying that in general a transfer of eukaryotic sequences into will not cause an efficient gene expression. However, there been reports on functionality 35S cauliflower mosaic virus (CaMV) bacteria. We therefore decided experimentally investigate capability plant direct expression various Accordingly, we tested ten different plant-specific promoters Solanum tuberosum, Nicotiana tabacum, CaMV, Agrobacterium tumefaciens, A. rhizogenes for ability initiate transcription five eubacterial species (Escherichia coli, Yersinia enterocolitica, Pseudomonas putida, Acinetobacter sp. BD413). To monitor strength bacteria created fusions between these coding region luciferase genes Vibrio harveyi measured luminescence Heterologous was observed 50% combinations analysed. then mapped start site caused by one promoters, ST-LS1 S. bacterial species. The location indicated themselves were recognised apparatus. recognition RNA polymerase further confirmed site-directed mutagenesis analysis effects mutations E. coli. Using mutants our reporter assays could localise serving as –10 results study show are much less specific than is generally assumed. This great importance knowledge about systems construction optimised vectors.