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Expression of Mycoplasma pulmonis antigens in Escherichia coli
1Department of Laboratory Animal Science, School of Hygienic Science, Kitasato University, Sagamihara, Japan.
Abstract:
The expression of Mycoplasma pulmonis antigen in Escherichia coli was investigated by cloning genomic DNA derived from M. pulmonis m 53, and the DNA fragment participating in antigen expression was identified. When the DNA library of M. pulmonis was screened by colony immunoassay using anti-M. pulmonis serum, 10 recombinant clones expressing seroreactive antigens were obtained. The recombinant plasmids isolated from these clones included 3.7-6.5 kilobase pair (kbp) DNA inserts, while all clones contained a common 2.3-kbp DNA fragment. Subcloning of initial DNA inserts showed that the common 2.3-kbp fragment is essential for antigen expression. Moreover, antiserum against the recombinant antigen generated from the 2.3-kbp DNA fragment recognized a native M. pulmonis antigen. The reactivity of this antiserum was absorbed specifically with M. pulmonis. These results suggest that the cloned 2.3-kbp DNA fragment codes an antigen specific to M. pulmonis.
Insights
Researchers identified a specific 2.3-kilobase pair DNA fragment from Mycoplasma pulmonis that is essential for expressing a key antigen. This finding advances understanding of Mycoplasma pulmonis antigen identification and characterization.
Area of Science:
- Microbiology
- Molecular Biology
- Immunology
Background:
- Mycoplasma pulmonis is a significant respiratory pathogen in rodents.
- Identifying specific antigens is crucial for developing diagnostic tools and vaccines.
Purpose of the Study:
- To clone and express Mycoplasma pulmonis antigen in Escherichia coli.
- To identify the specific DNA fragment responsible for antigen expression.
Main Methods:
- Construction of a genomic DNA library from Mycoplasma pulmonis m 53 in E. coli.
- Screening of the DNA library using colony immunoassay with anti-Mycoplasma pulmonis serum.
- Subcloning of DNA inserts to pinpoint the essential fragment for antigen expression.
Main Results:
- Ten recombinant clones expressing seroreactive antigens were identified.
- A common 2.3-kilobase pair (kbp) DNA fragment was found in all expressing clones and confirmed essential for antigen expression.
- Antiserum against the recombinant antigen recognized native Mycoplasma pulmonis antigen and was specifically absorbed by Mycoplasma pulmonis.
Conclusions:
- The cloned 2.3-kbp DNA fragment codes for an antigen specific to Mycoplasma pulmonis.
- This fragment is a promising target for developing diagnostic and potentially therapeutic strategies against Mycoplasma pulmonis infections.