Guzmán C A, Walker M J, Rohde M, Timmis K N
Department of Microbiology, GBF-National Research Center for Biotechnology, Braunschweig, Germany.
Infect Immun. 1991 Oct;59(10):3787-95. doi: 10.1128/iai.59.10.3787-3795.1991.
Nonfused (i.e., nonhybrid) filamentous hemagglutinin (FHA) of Bordetella pertussis was efficiently expressed in Escherichia coli K-12 and Salmonella typhimurium aroA at levels higher than those found in wild-type B. pertussis when the upstream signals of the gene were replaced and the translation initiation region was engineered to optimize translational efficiency. Inclusion of part of the C-terminal FHA open reading frame, whose translation product does not appear to be part of the major secreted species of FHA, was shown to be important in achieving protein expression in both E. coli and S. typhimurium aroA; removal of the downstream gene sequence abolished recombinant FHA production. The levels of expression observed varied widely according to the construct and host bacterium used.
当百日咳博德特氏菌基因的上游信号被替换且翻译起始区域经改造以优化翻译效率时，百日咳博德特氏菌的未融合（即非杂交）丝状血凝素（FHA）在大肠杆菌K-12和鼠伤寒沙门氏菌aroA中高效表达，表达水平高于野生型百日咳博德特氏菌中的水平。包含部分C端FHA开放阅读框（其翻译产物似乎不是FHA主要分泌形式的一部分）被证明对于在大肠杆菌和鼠伤寒沙门氏菌aroA中实现蛋白表达很重要；去除下游基因序列则消除了重组FHA的产生。根据所使用的构建体和宿主细菌的不同，观察到的表达水平差异很大。
