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A broad-host-range vector of incompatibility group Q can work as a plasmid vector in Neisseria meningitidis: a new genetical tool

Department of Bacteriology, National Institute of Infectious Diseases, Toyama 1-23-1, Shinjuku-ku, Tokyo 162-8640, Japan¹

Author for correspondence: Hideyuki Takahashi. Tel: +81 3 5285 1111. Fax: +81 3 5285 1163. e-mail: hideyuki{at}nih.go.jp

Plasmid pHT128, a derivative of the broad-host-range IncQ vector pGSS33, was successfully introduced into Neisseria meningitidis. Under optimal conditions, pHT128 was transferred from Escherichia coli to N. meningitidis by triparental conjugation at a frequency of 10^-5–10^-6. The copy number of pHT128 in N. meningitidis was almost the same as in E. coli, in which the copy number of IncQ plasmids per chromosome is estimated to be 10. pHT128 was maintained as an episome in N. meningitidis in the presence of chloramphenicol, a marker of the plasmid. It was also shown that an opc or pilE1 gene cloned on pHT128 could be expressed in N. meningitidis under control of the tac promoter and could complement a mutation of opc or pilE1, respectively. In addition, the conjugational introduction of pHT128 into N. meningitidis was demonstrated to be independent of natural transformation competence. All the results indicate that pHT128 is a useful vector for N. meningitidis as a new genetical tool.

Keywords: IncQ plasmid, triparental conjugation, restriction/modification systems, complementation

Abbreviations: Amp^r/Amp^s, ampicillin resistant/sensitive; Cml^r, chloramphenicol resistant; Erm^r, erythromycin resistant; Kan^r/Kan^s, kanamycin resistant/sensitive; Tet^r/Tet^s, tetracycline resistant/sensitive; Str^r/Str^s, streptomycin resistant/sensitive

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