Cloning and expression of the Legionella micdadei "common antigen" in Escherichia coli

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To study individual Legionella antigens, a Legionella micdadei genomic library in Escherichia coli SC181 was established. Partially Sau3A digested L. micdadei DNA fragments (15-25 kilobase pairs (kb] were cloned into the tetracycline resistance gene of the cosmid vector pHC79. Four thousand ampicillin resistant recombinants were obtained; seven hundred were screened for expression of Legionella antigens in Western blot analysis with a polyspecific E. coli-absorbed anti-L. micdadei rabbit antibody. One of the positive clones expressed a 60 kilodalton (K) antigen, which reacted strongly with a monospecific rabbit antiserum raised against L. micdadei "common antigen" (CA), and an additional 13 K L. micdadei protein. The region encoding these two proteins from the 17 kb recombinant plasmid (pBA 2) was subcloned in pBGS18+. The DNA sequence of the CA encoding region in the 2.7 kb subcloned fragment will provide important information with respect to genetic vs. antigenic relatedness among Legionellae and other Gram-negative species, as well as to CA structure and possible function.
Original languageEnglish
JournalA P M I S. Acta Pathologica, Microbiologica et Immunologica Scandinavica
Volume97
Issue number1
Pages (from-to)14-22
Number of pages9
ISSN0903-4641
Publication statusPublished - 1989

    Research areas

  • Antigens, Bacterial, Bacterial Proteins, Cloning, Molecular, Escherichia coli, Immunoelectrophoresis, Two-Dimensional, Legionella, Molecular Weight, Restriction Mapping

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