National Institute of Animal Health (NIAH)

Topics in Animal Health Research 2013

05. Development of a gene expression vector and a transformation protocol for Melissococcus plutonius

Japanese

Melissococcus plutonius is an important pathogen that causes European foulbrood in honey bee larvae. Despite its importance, our understanding of this bacterium remains very limited. A lack of tools for molecular approaches was a factor that hampered studies on M. plutonius. Therefore, in this study, we constructed a novel gene expression vector for M. plutonius and designed an electroporation protocol to introduce the vector into M. plutonius strains. The developed vector pMX2 consists of a broad-host-range replication origin, the Streptococcus suis malX promoter, multiple cloning sites, and the chloramphenicol-resistance gene. pMX2 can be introduced into M. plutonius strain ATCC 35311 by the designed electroporation protocol at an average efficiency of 1.76 × 103 transformants/μg DNA. Because the S. suis malX promoter works in M. plutonius too, even promoterless genes can be expressed and analyzed in M. plutonius using the promoter. Therefore, these genetic tools should promote future research on M. plutonius.
(Bacterial and Parasitic Disease Research Division)

References:

Takamatsu D. et al. (2013) Appl. Environ. Microbiol. 79:3882-3886

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