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2012 | 59 | 2 |
Tytuł artykułu

High efficiency method to obtain supercoiled DNA with a commercial plasmid purification kit

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Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
Supercoiled state corresponds to the active form for plasmid applications. The relaxed circular form of plasmids is often inactive or poorly active. To obtain significant amounts of almost fully supercoiled DNA, we modified the standard protocol of a commercially; available Qiagen plasmid purification kit. Our changes led to isolation of almost 100% of the plasmids in the supercoiled state. The modified protocol was used to purify different plasmids with consistent results. The purified plasmids maintain supercoiled state for about two months. The modified protocol is very advantageous because it allows easy DNA production with high degree of supercoiled form at low cost.
Słowa kluczowe
Wydawca
-
Rocznik
Tom
59
Numer
2
Opis fizyczny
p.275-278,fig.,ref.
Twórcy
autor
  • Department of Structural and Functional Biology, University of Naples Federico II, Naples, Italy
autor
autor
autor
Bibliografia
  • Anderson RJ, Schneider J (2007) Plasmid DNA and viral vector-based vaccines for the treatment of cancer. Vaccine 25 (Suppl 2): B24-B34.  
  • Cupillard L, Juillard V, Latour S, Colombet G, Cachet N, Richard S, Blanchard S, Fischer L (2005) Impact of plasmid supercoiling on the efficacy of a rabies DNA vaccine to protect cats. Vaccine 23: 1910-1916. 
  • Diogo MM, Queiroz JA, Prazeres DM (2005) Chromatography of plasmid DNA. J Chromatogr A 1069: 3-22. 
  • Doyle PS, Ladoux B, Viovy JL (2000) Dynamics of a tethered polymer in shear flow. Phys Rev Lett 84: 4769-4772. 
  • Hoffman MM, Khrapov MA, Cox JC, Yao J, Tong L, Ellington AD (2004) AANT: the Amino Acid-Nucleotide Interaction Database. Nucleic Acids Res 32: D174-D181. 
  • Latulippe DR, Zydney AL (2011) Separation of plasmid DNA isoforms by highly converging flow through small membrane pores. J Colloid Interface Sci 357: 548-553. 
  • Lee MP, Sander M, Hsieh TS (1989) Single strand DNA cleavage reaction of duplex DNA by Drosophila Topoisomerase II. J Biol Chem 264: 13510-1358. 
  • Luscombe NM, Laskowski RA, Thornton JM (2001) Amino acid-base interactions: a three-dimensional analysis of protein-DNA interactions at an atomic level. Nucleic Acids Res 29: 2860-2874. 
  • Osheroff N, Shelton ER, Brutlag DL (1983) DNA topoisomerase II from Drosophila melanogaster. Relaxation of supercoiled DNA. J Biol Chem 258: 9536-9543. 
  • Prazeres DM, Schluep T, Cooney C (1998) Preparative purification of supercoiled plasmid DNA using anionexchange chromatography. J Chromatogr A 806: 31-45. 
  • Sousa A, Sousa F, Queiroz JA (2009) Biorecognition of supercoiled plasmid DNA isoform in lysine-affinity chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 877: 3257-3260.  
  • Sousa A, Sousa F, Queiroz JA (2010) Differential interactions of plasmid DNA, RNA and genomic DNA with amino acid-based affinity matrices. J Sep Sci 33: 2610-2618. 
  • Sousa F, Prazeres DM, Queiroz JA (2008) Affinity chromatography approaches to overcome the challenges of purifying plasmid DNA. Trends Biotechnol 26: 518-525. 
  • Sousa F, Tomaz CT, Prazeres DM, Queiroz JA (2005) Separation of supercoiled and open circular plasmid DNA isoforms by chromatography with a histidine-agarose support. Anal Biochem 343: 183-15. 
  • Stadler J, Lemmens R, Nyhammar T (2004) Plasmid DNA purification. J Gene Med 6: S54-S66
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PL
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Typ dokumentu
Bibliografia
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