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Insertion into the Mycobacterium smegmatis genome of the aph gene through lysogenization with the temperate
E Anes1, I Portugal, J Moniz-Pereira
1Laboratório de Microbiologia Molecular, Faculdade de Farmácia, Universidade de Lisboa, Portugal.
Abstract:
A derivative of the temperate mycobacteriophage Ms6 containing the aph gene from transposon Tn5 was constructed. In the transductants the aph gene was integrated in the bacterial genome. The aph gene is stably maintained in the absence of positive selection after more than 150 generations. The results presented in this report show that Ms6 can be used as a vehicle for the integration of foreign DNA into the Mycobacterium smegmatis genome.
Insights
Researchers engineered the mycobacteriophage Ms6 to carry the aph gene. This gene integrates stably into the Mycobacterium smegmatis genome, demonstrating Ms6
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Mycobacteriophage Ms6 is a temperate phage.
- Transposon Tn5 carries the aph gene.
- Foreign DNA integration into mycobacteria is challenging.
Purpose of the Study:
- To construct a derivative of mycobacteriophage Ms6.
- To evaluate its efficacy as a vehicle for foreign DNA integration into Mycobacterium smegmatis.
Main Methods:
- Construction of a recombinant Ms6 phage carrying the aph gene.
- Transduction of Mycobacterium smegmatis with the engineered phage.
- Assessment of aph gene stability in the bacterial genome over generations.
Main Results:
- The aph gene was successfully integrated into the Mycobacterium smegmatis genome.
- The integrated aph gene demonstrated stable maintenance.
- Stability was observed for over 150 generations without positive selection.
Conclusions:
- Mycobacteriophage Ms6 serves as an effective vehicle for integrating foreign DNA.
- The engineered Ms6 derivative facilitates stable genetic modification of Mycobacterium smegmatis.