Plasmid of DNA

Hansma H G, Vesenka J, Siegerist C, Kelderman G, Morrett H, Sinsheimer R L, Bustamante C, Elings V and Hansma P K 1992 Reproducible imaging and dissection of plasmid DNA under liquid with the atomic force microscope Science 256 1180... 

Most plasmids are topologically closed circles of DNA. They can be separated from the bulk of the chromosomal DNA by virtue of their resistance to alkaline solution. The double-stranded stmcture of DNA is denatured at high pH, but because the two strands of the plasmid are topologically joined they are more readily renatured. This property is exploited in rapid procedures for the isolation of plasmid DNA from recombinant microorganisms (5,6). 

Plasmid Vectors for Facile Introduction of Passenger DNA and Selection of Recombinants. The map of a commonly used plasmid vector, pUC19 (7), is shown in Figure 2. Three parts of the vector are key to its utility. The origin sequence, oh, allows the repHcation of plasmid DNA in high copy number relative to the chromosome. A gene, amp, encoding the enzyme beta-lactamase, which hydrolyzes penicillin compounds, allows... 

Toyopearl HW-75 resin, with pores larger than 1000 A, have been used in place of ultracentrifugation steps for the purification of plasmid DNA. Ultracentrifugation is a time-consuming process and requires expensive chemicals, such as cesium chloride. Toyopearl HW-75 resin provides superior separation performance for plasmid DNA and also provides high yields (54). 

Continuous. Long fermentation run times, with many generations, increase the chance of mutation or loss of plasmid DNA. 

Liu MA, Ulmer JB (2005) Human clinical trials of plasmid DNA vaccines. Adv Genet 55 25-40... 

Griffin DE III, Hill WE. 1978. In vitro breakage of plasmid DNA by mutagens and pesticides. Mutat Res 52 161-169. 

S., Hamer, )., Escobedo, J., Cohen, F., Radhakrishnan, R., Dwarki, V, and ZuCKERMANN, R.N. Lipitoids — novel cationic lipids for cellular delivery of plasmid DNA in vitro. Chem. Biol. 1998, 5, 345-354. 

FIG. 2 Atomic force microscopy of plasmid DNA at different stages of condensation with polycations. (a) Circular plasmid DNA (b) DNA condensed with poly-L-lysine (PLL, mol. wt. 4 kDa) at a DNA phosphate/lysine ratio of 2 1 (c) DNA condensed with PLL (mol. wt. 10 kDa) at a phos-phate/lysine ratio of 1 2 (d) toroid of DNA condensed with PLL-asialo-orosomucoid conjugate at a phosphate/lysine ratio of 1 6. [(a) and (d) from Ref. 80, copyright 1998 Oxford University Press (b) and (c) reprinted with permission from Ref. 66, copyright 1999 American Chemical Society.]... 

Fig. 3. Comparison of transfection efficiencies obtained using PolyFect Reagent, a dendrimer-based transfection reagent, and a calcium phosphate-mediated procedure. COS-7 and HeLa cells were transfected in srx-weU plates with a /3-galactosidase expression plasmid using the appropriate protocol. For the calcium phosphate-mediated transfection, 6 pg of plasmid DNA was used and the medium was changed after 5 h incubation. Transfections were performed in triplicate, and transfection efficiency was measured by monitoring the /3-galactosidase activity of extracts obtained from the transfected cells. The amoimt of /3-galactosidase activity in the extracts correlates with the transfection efficiency. Cells were harvested 48 h post-trans-fection...

Isolation of plasmid DNA, transformation, transfection and cloning procedures were performed using standard techniques (Sambrook et ut/., 1989). 

In the case of plasmid DNA the intracellular target (i.e., transcription machinery) is located in the nucleus. Plasmid DNA is a much larger molecule than oligonucleotides, and... 

M. Nishikawa, S. Takamura, F. Yamashita, Y. Takahura, D. K. Meijer, M. Hashida, and J. P. Swart, Pharmacokinetics and in vivo gene transfer of plasmid DNA complexed with mannosylated poly(L-lysine) in mice, J. Drug Target., 8 (2000) 29-38. 

Blessing, T., Remy, J.S., and Behr, J.P., Monomolecular collapse of plasmid DNA into stable virus-like particles, Proceedings of the National Academy of Sciences, United States of America, 1998, 95, 1427-1431. 

Meyer M, Wagner E (2006) Recent developments in the application of plasmid DNA-based vectors and small interfering RNA therapeutics for cancer. Hum Gene Ther 17 1062-1076... 

Nishikawa M, Yamauchi M, Morimoto K, Ishida E, Takakura Y, Hashida M (2000) Hepato-cyte-targeted in vivo gene expression by intravenous injection of plasmid DNA complexed with synthetic multi- functional gene delivery system. Gene Ther 7 548-555... 

Saito G, Amidon GL, Lee KD (2003) Enhanced cytosolic delivery of plasmid DNA by a sulfhydryl-activatable listeriolysin O/protamine conjugate utilizing cellular reducing potential. Gene Ther 10 72-83... 

Lechardeur D, Verkman AS, Lukacs GL (2005) Intracellular routing of plasmid DNA during non-viral gene transfer. Adv Drug Deliv Rev 57 755-767... 

Lechardeur D, Lukacs GL (2006) Nucleocytoplasmic transport of plasmid DNA a perilous journey from the cytoplasm to the nucleus. Hum Gene Ther 17 882-889... 

Nagasaki T, Myohoji T, Tachibana T, Futaki S, Tamagaki S (2003) Can nuclear localization signals enhance nuclear localization of plasmid DNA Bioconjug Chem 14 282-286... 

Interaction of polyamine gene vectors with RNA leads to the dissociation of plasmid DNA-carrier complexes. J Gene Med 8 1416-1424... 

Itaka K, Harada A, Yamasaki Y, Nakamura K, Kawaguchi H, Kataoka K (2004) In situ single cell observation by fluorescence resonance energy transfer reveals fast intra-cytoplasmic delivery and easy release of plasmid DNA complexed with linear polyethylenimine. J Gene Med 6 76-84... 

Matsumoto Y, Itaka K, Yamasoba T, Kataoka K (2009) Intranuclear fluorescence resonance energy transfer analysis of plasmid DNA decondensation from nonviral gene carriers. J Gene Med 11 615-623... 

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