Walker M J, Guzmán C A, Rohde M, Timmis K N
Department of Microbiology, GBF-National Research Centre for Biotechnology, Braunschweig, Federal Republic of Germany.
Infect Immun. 1991 May;59(5):1739-46. doi: 10.1128/iai.59.5.1739-1746.1991.
Serotype-specific fimbriae of Bordetella pertussis are considered potential components of new-generation vaccines against whooping cough. Attempts to characterize fimbriae, and indeed other virulence determinants, produced by B. pertussis have been frustrated on one hand by low yields from B. pertussis itself and on the other by an inability to produce native recombinant products in Escherichia coli. In order to try to circumvent this problem, we have examined the expression of B. pertussis serotype 2 fimbriae in Bordetella parapertussis and Bordetella bronchiseptica from native as well as E. coli expression signals. These studies revealed that the fimbrial gene product was expressed from the original B. pertussis promoter and Shine-Dalgarno sequence in both B. parapertussis and B. bronchiseptica. The transcriptional start site of the gene was located 146 nucleotides upstream of its ATG start codon. A recombinant fimbrial subunit gene containing PLAC and the atpE translation initiation region of E. coli was also expressed in B. bronchiseptica. In all cases in which gene expression was detected the gene product was expressed as serotype 2-specific fimbriae as determined by enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopic investigation of the bacterial cell surface.
百日咳博德特氏菌的血清型特异性菌毛被认为是新一代百日咳疫苗的潜在组成部分。一方面，由于百日咳博德特氏菌本身产量较低，另一方面由于无法在大肠杆菌中产生天然重组产物，对百日咳博德特氏菌产生的菌毛以及其他毒力决定因素进行表征的尝试一直受挫。为了试图解决这个问题，我们研究了来自天然以及大肠杆菌表达信号的百日咳博德特氏菌血清型2菌毛在副百日咳博德特氏菌和支气管败血博德特氏菌中的表达。这些研究表明，菌毛基因产物在副百日咳博德特氏菌和支气管败血博德特氏菌中均从原始百日咳博德特氏菌启动子和夏因-达尔加诺序列表达。该基因的转录起始位点位于其ATG起始密码子上游146个核苷酸处。含有大肠杆菌PLAC和atpE翻译起始区域的重组菌毛亚基基因也在支气管败血博德特氏菌中表达。在所有检测到基因表达的情况下，通过酶联免疫吸附测定（ELISA）和对细菌细胞表面的免疫电子显微镜研究确定，基因产物均表达为血清型2特异性菌毛。
