Related Experiment Videos
Molecular properties of the erythromycin resistance plasmid pPV141 from Staphylococcus chromogenes
G A Somkuti1, D K Solaiman, D H Steinberg
1U.S. Department of Agriculture, Eastern Regional Research Center, Wyndmoor, Pennsylvania 19038, USA.
Abstract:
The 2.3-kb erythromycin resistance (EmR) plasmid pPV141 of Staphylococcus chromogenes 3688 was isolated and characterized. Nucleotide sequence analysis identified ORF1 and ORF2 separated by a 445-bp spacing, encoding a 158-residue replication protein (Rep141) and a 244-residue erythromycin resistance protein (Erm, rRNA adenine N-6-methyltransferase), respectively. Structural analysis and Southern hybridization showed that the rep and ermM genes in pPV141 shared homology with other known EmR plasmids. Based on sequence analysis, pPV141 was classified as a unique member of the pSN2 family of EmR plasmids.
Insights
The Staphylococcus chromogenes plasmid pPV141 confers erythromycin resistance (EmR). This unique plasmid, belonging to the pSN2 family, contains genes for replication and EmR, showing homology to other resistance plasmids.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Erythromycin resistance (EmR) is a significant concern in Staphylococcus species.
- Plasmids are key vectors for antibiotic resistance gene dissemination.
- Characterization of novel resistance plasmids aids in understanding bacterial evolution.
Purpose of the Study:
- To isolate and characterize the erythromycin resistance plasmid pPV141 from Staphylococcus chromogenes 3688.
- To determine the genetic elements responsible for erythromycin resistance and plasmid replication.
- To classify pPV141 within known plasmid families.
Main Methods:
- Plasmid isolation and purification from S. chromogenes.
- Nucleotide sequencing and open reading frame (ORF) identification.
- Southern hybridization for gene homology analysis.
- Bioinformatic analysis for gene and protein structure prediction.
Main Results:
- The 2.3-kb plasmid pPV141 was successfully isolated and sequenced.
- Two ORFs were identified: ORF1 encoding a replication protein (Rep141) and ORF2 encoding an erythromycin resistance protein (Erm).
- The rep and ermM genes showed homology to those in other EmR plasmids.
- Sequence analysis classified pPV141 as a unique member of the pSN2 family.
Conclusions:
- Plasmid pPV141 is a novel EmR plasmid in Staphylococcus chromogenes.
- The identified genes (Rep141 and Erm) are responsible for plasmid replication and erythromycin resistance.
- pPV141 represents a unique variant within the pSN2 family of EmR plasmids, contributing to the understanding of antibiotic resistance mechanisms.