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Regions of Mycoplasma pneumoniae cytadhesin P1 structural gene exist as multiple copies
C J Su1, A Chavoya, J B Baseman
1Department of Microbiology, University of Texas Health Science Center, San Antonio 78284-7758.
Abstract:
The Mycoplasma pneumoniae cytadhesin P1 structural gene with flanking regions was labeled by nick translation and used as a probe to analyze gene copy number in M. pneumoniae. Multiple bands of genomic DNA were hybridized by the probe. To establish what part of the P1 gene existed as multiple copies, the P1 gene and regions adjacent to the 3' and 5' ends were divided with restriction enzymes into 14 segments ranging in size from 174 to 651 base pairs. These pieces were purified on agarose gels, subcloned into pUC19, purified, labeled by nick translation, and used to probe the entire M. pneumoniae genome. Several regions near the middle and carboxy end of the P1 structural gene hybridized to single copies. The remaining P1 subclones hybridized to multiple bands under stringent hybridization conditions, indicating extensive homology with other parts of the M. pneumoniae genome. The single- versus multiple-copy nature of P1 structural gene domains is discussed.
Insights
Mycoplasma pneumoniae
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Mycoplasma pneumoniae is a significant human respiratory pathogen.
- The cytadhesin P1 protein is crucial for M. pneumoniae adherence to host cells.
- Understanding the P1 gene's structure and copy number is key to studying its regulation.
Purpose of the Study:
- To investigate the gene copy number of the Mycoplasma pneumoniae cytadhesin P1 structural gene.
- To determine which regions of the P1 gene exist as single or multiple copies within the M. pneumoniae genome.
Main Methods:
- Nick translation labeling of the P1 gene and flanking regions.
- Restriction enzyme digestion and agarose gel purification of P1 gene segments.
- Subcloning into pUC19, purification, and nick translation labeling of segments.
- Hybridization of labeled P1 gene segments to the M. pneumoniae genome under stringent conditions.
Main Results:
- The P1 gene probe hybridized to multiple bands of M. pneumoniae genomic DNA.
- Specific regions near the middle and carboxy terminus of the P1 gene hybridized to single copies.
- Other P1 gene segments hybridized to multiple bands, indicating extensive homology with other genomic regions.
Conclusions:
- The Mycoplasma pneumoniae P1 gene exhibits both single- and multiple-copy domains.
- Extensive homology exists between parts of the P1 gene and other regions of the M. pneumoniae genome.
- This suggests potential complexity in P1 gene expression and regulation.