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Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...

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Phage-Mediated Genetic Manipulation of the Lyme Disease Spirochete Borrelia burgdorferi

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Transformation of Anaplasma marginale.

Roderick F Felsheim¹, Adela S Oliva Chávez, Guy H Palmer

¹Department of Entomology, University of Minnesota, St. Paul, MN 55108, USA.

Veterinary Parasitology

|October 20, 2009

Summary

Researchers successfully genetically modified Anaplasma marginale, a tick-borne pathogen causing bovine anaplasmosis. This breakthrough enables future studies on this economically significant cattle disease.

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Forward Genetic Approaches in Chlamydia trachomatis

Forward Genetic Approaches in Chlamydia trachomatis

Published on: October 23, 2013

Area of Science:

Microbiology
Genetics
Veterinary Science

Background:

Anaplasma marginale causes bovine anaplasmosis, a disease with substantial global economic impact on cattle farming.
The obligate intracellular nature of A. marginale hinders genetic manipulation, a common challenge for related prokaryotes like Ehrlichia and Rickettsia.

Purpose of the Study:

To develop a method for genetically transforming Anaplasma marginale.
To create plasmid constructs for introducing TurboGFP and antibiotic resistance genes into A. marginale.

Main Methods:

Co-electroporation of transposon and transposase expression plasmids into A. marginale.
Selection of transformed bacteria using spectinomycin and streptomycin.
Analysis of gene insertion sites via cloning and DNA sequencing.

Main Results:

Successfully generated fluorescent Anaplasma marginale transformants expressing TurboGFP and antibiotic resistance genes.
Demonstrated insertion of the transposon into the native tr region of A. marginale via homologous recombination, not transposase activity.
Observed that transformants exhibit slower growth and require longer subculture intervals compared to wild-type.

Conclusions:

A. marginale can be genetically transformed using a homologous recombination strategy.
This transformation method holds potential for other Anaplasma and Ehrlichia species.
The developed technique facilitates future research into A. marginale biology and disease control.