Abstract
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 language | English |
|---|---|
| Journal | A P M I S. Acta Pathologica, Microbiologica et Immunologica Scandinavica |
| Volume | 97 |
| Issue number | 1 |
| Pages (from-to) | 14-22 |
| Number of pages | 9 |
| ISSN | 0903-4641 |
| Publication status | Published - 1989 |
Keywords
- Antigens, Bacterial
- Bacterial Proteins
- Cloning, Molecular
- Escherichia coli
- Immunoelectrophoresis, Two-Dimensional
- Legionella
- Molecular Weight
- Restriction Mapping