Abstract
Bordetella pertussis UT25 DNA was cloned into the kanamycin resistance gene of cosmid pCP13 to construct a genomic library in Escherichia coli LE392. One clone containing plasmid pDB441 expressed the filamentous hemagglutinin (FHA) as identified by protein immunoblots with the use of rabbit anti-B. pertussis antiserum, rabbit anti-FHA antiserum, and a monoclonal antibody to FHA. FHA is a protein of 220 to 210 kilodaltons, but the immunoreactive FHA, as expressed in E. coli, was larger than that expressed in B. pertussis, suggesting that there was a difference in the processing of this protein between these two bacteria. The fha gene was mapped to a 6.5-kilobase pair DNA fragment by the use of various restriction endonucleases. The kanamycin resistance gene of pCP13 was found to provide the promoter function but probably not the translation start signal for the fha gene. Conjugative transfer of pDB441 to B. pertussis BP353, a transposon Tn5-induced FHA mutant, increased the expression of the FHA over that seen with wild-type B. pertussis.
Publisher
American Society for Microbiology
Subject
Infectious Diseases,Immunology,Microbiology,Parasitology
Reference47 articles.
1. Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn 9;Alton N. K.;Nature (London),1979
2. Separation and characterization of two distinct hemagglutinins contained in purified leukocytosispromoting factor from Bordetella pertussis;Arai H.;Biochim. Biophys. Acta,1976
3. Insertion, excision, and inversion of Tn 5;Berg D. E.;Cold Spring Harbor Symp. Quant. Biol.,1981
4. L'endotoxine coquelucheuse;Bordet J.;Ann. Inst. Pasteur (Paris),1909
5. Cowell J. L. Y. Sato H. Sato B. An der Lan and C. R. Manclark. 1982. Separation purification and properties of the filamentous hemagglutinin and the leukocytosis promoting factor-hemagglutinin from Bordetella pertussis p. 371-379. In J. B. Robbins J. C. Hill and J. C. Sadoff (ed.) Bacterial vaccines. Thieme-Stratton Inc. New York.
Cited by
29 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献