| Autor: |
Suhan ML; Department of Microbiology and Immunology, Robert E. Byrd Health Sciences Center, West Virginia University, P.O. box 9177, Morgantown, WV 26506, USA., Thompson HA |
| Jazyk: |
angličtina |
| Zdroj: |
FEMS microbiology letters [FEMS Microbiol Lett] 2000 Mar 15; Vol. 184 (2), pp. 303-6. |
| DOI: |
10.1111/j.1574-6968.2000.tb09031.x |
| Abstrakt: |
Coxiella burnetii can be transformed to ampicillin resistance by electroporation with plasmids encoding beta-lactamase. However, non-plasmid emergence of resistance to ampicillin also develops. To validate the usefulness of the bla gene marker for selection and detection, transformed C. burnetii were examined for beta-lactamase expression by use of immunoblotting after SDS-PAGE. The 29-kDa mature form of the beta-lactamase protein was detected in C. burnetii lysates. Quantitation of these immunoblot signals showed that C. burnetii surprisingly expressed low levels of beta-lactamase. The results validate the use of plasmid-encoded ampicillin resistance as a means for selecting C. burnetii transformants; they also suggest that levels of ampicillin used for selection pressure should be empirically determined and that detection of beta-lactamase by antibody blotting done to confirm transformants. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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