Development and applications of a DNA labeling method  with magnetic nanoparticles to study the role of horizontal gene  transfer events between bacteria in soil pollutant  bioremediation processes.
Horizontal gene transfers are critical mechanisms of bacterial evolution and adaptation that are involved to a significant level in the degradation of toxic molecules such as xenobiotic pesticides. However, understanding how these mechanisms are regulated in situ and how they could be used by man to increase the degradation potential of soil microbes is compromised by conceptual and technical limitations. This includes the physical and chemical complexity and heterogeneity in such environments leading to an extreme bacterial taxonomical diversity and a strong redundancy of genes and functions. In addition, more than 99 % of soil bacteria fail to develop colonies in vitro, and even new DNA-based investigation methods (metagenomics) are not specific and sensitive enough to consider lysis recalcitrant bacteria and those belonging to the rare biosphere. The objective of the ANR funded project “Emergent” was to develop a new culture independent approach to monitor gene transfer among soil bacteria by labeling plasmid DNA with magnetic nanoparticles in order to specifically capture and isolate recombinant cells using magnetic microfluidic devices. We showed the feasibility of the approach by using electrotransformation to transform a suspension of Escherichia coli cells with biotin-functionalized plasmid DNA molecules linked to streptavidin-coated superparamagnetic nanoparticles. Our results have demonstrated that magnetically labeled cells could be specifically retained on micromagnets integrated in a microfluidic channel and that an efficient selective separation can be achieved with the microfluidic device. Altogether, the project offers a promising alternative to traditional culture-based approaches for deciphering the extent of horizontal gene transfer events mediated by electro or natural genetic transformation mechanisms in complex environments such as soil.
J
J
Pivetal
M
M
Frénéa-Robin
N
N
Haddour
C
C
Vézy
L F
LF
Zanini
G
G
Ciuta
N M
NM
Dempsey
F
F
Dumas-Bouchiat
G
G
Reyne
S
S
Bégin-Colin
D
D
Felder-Flesh
C
C
Ghobril
G
G
Pourroy
P
P
Simonet
D001419:Bacteria / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D001673:Biodegradation, Environmental; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D004867:Equipment Design; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000254:growth & development; D005602:France; D022761:Gene Transfer, Horizontal; D058185:Magnetite Nanoparticles / Q000737:chemistry*; D044085:Microfluidics; D010957:Plasmids; D012989:Soil Pollutants / Q000032:analysis*
D058185:Magnetite Nanoparticles; D012989:Soil Pollutants; D004247:DNA