Gold and Silver Nanoparticles – Effective Agents for Elimination the Plasmids of Antibiotic Resistance

Dybkova S. M.

NANOTECHNOLOGY

Scentific article

Unique properties of metal nanoparticles and their biological activity open wide perspectives for their using in medicine. Studies of the peculiarities of metal’s nanoparticles interaction with nucleic acids of plasmids has fundamental importance in the investigation of metal’s nanoparticles influence on genetic complex of cells. The research has been performed with using model systems of nucleic acids such as plasmids pUC19 and pBR322. These plasmids (R­plasmids) have genes of resistance to antibiotics: plasmid pUC19 have gene Ampr (GenBank Assession Number LO9137); plasmid pBR322 have gene Ampr and Tetr (GenBank Assession Number J01749). The plasmids pUC19 and pBR322 were living in bacterias of strain E. coli XL1­Blue Kan (Stratagene). Plasmids obtained by method of Birnboim&Doly and stored at – 200с. This plasmids were influenced by gold and silver nanoparticles in different concentrations: gold nanoparticles 30 nm – 9,6 and 3,8 mkg/ml, silver nanoparticles 30 nm – 43,0 and 17,0 mkg/ml. By electrophoresis method it has been shown the changes in electrophoretic mobility of plasmids pUC19 and pBR322. These results indicate the changes in the structure of the model plasmids. Interaction of gold and silver nanoparticles of different size with plasmids pUC19 and pBR322 has been per­ formed by the method of electron microscopy. Nanoparticles concentation in incubation mixtures: gold nanopar­ ticles 20 nm – 5,8 mkg/ml, 30 nm – 9,6 mkg/ml; silver nanoparticles 30 нм – 43,0 and 17,0 mkg/ml. Gold nanoparticles (20 and 30 nm) causing relaxation of plasmids placed not sorted on it’s threads. Moreover, gold nanoparticles with size 20 nm, in contrast to the 30 nm nanoparticles, aggregated an untwisted plasmids. Silver nanoparticles with size 30 nm leads to destructive changes of plasmid DNA, while being located on it at regu­ lar intervals. Thus, interaction of gold (20 and 30 nm) and silver nanoparticles (30 nm) with plasmids pUC19 and pBR322 resulting in plasmid’s relaxation. Elimination of the plasmids pUC19 and pBR322 was performed by culturing the bacteria E. coli XL1­Blue (pUC19) and E. coli XL1­Blue (pBR322) with gold and silver nanoparticles within 18 hours. Gold (20 and 30 nm) and silver (30 nm) nanoparticles cause a high frequency of plasmid’s pUC19 and pBR322 elimination. In processing the plasmids of bacteria E. coli XL1­Blue (pUC19) and E. coli XL1­Blue (pBR322) gold nanoparticles of 20 nm at a concentration of 5. 6 mg / ml elimination rate was 89 % and 86 % respectively. In the case of gold nanoparticles of 30 nm (9. 6 mg/ ml) rate of elimination was: pUC19 – 91 %, and pBR322 – 90 %. Silver nanoparticles at concentration of 17 mg/ml eliminate plasmids from strains of E. coli XL1­Blue (pUC19) and E. coli XL1­Blue (pBR322) with frequency: pUC19 – 98 % and pBR322 – 96 %. Frequency of spontaneous elimination was: in the case of plasmid pUC19 – 37 %, pBR322 – 34 %. Thus, effectively eliminates of plasmid’s pUC19 and pBR322 has been performed by gold (20 and 30 nm) and silver (30 nm) nanoparticles. For a final conclusion about gold and silver nanoparticles’ properties to effectively elimination of R­plasmids to need hold in further studies involving the plasmids of clinical isolates of bacterias

gold nanoparticles; silver nanoparticles; plasmids; DNA; elimination

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«Bulletin of problems biology and medicine» Issue 3 part 2 (111), 2014 year, 314-318 pages, index UDK 577. 352. 3:616­006. 04:57. 02:661. 859