Integration of Bacteria Capture via Filtration and in Situ Lysis for Recovery of Plasmid DNA under Industry-Compatible Conditions

2007 ◽  
Vol 23 (4) ◽  
pp. 895-903 ◽  
Author(s):  
Kevin O'Mahony ◽  
Ruth Freitag ◽  
Frank Hilbrig ◽  
Ivo Schumacher ◽  
Patrick Müller
Keyword(s):  
Plasmid Dna

In situ gelling and mucoadhesive polymer vehicles for controlled intranasal delivery of plasmid DNA

2001 ◽  
Vol 59 (1) ◽  
pp. 144-151 ◽  
Author(s):  
Jeong-Sook Park ◽  
Yu-Kyoung Oh ◽  
Ho Yoon ◽  
Jung Mogg Kim ◽  
Chong-Kook Kim
Keyword(s):  
Plasmid Dna ◽  
Intranasal Delivery ◽  
Mucoadhesive Polymer ◽  
In Situ Gelling

Effect of ozone on plasmid DNA of Escherichia coli in situ

1987 ◽  
Vol 21 (7) ◽  
pp. 823-827 ◽  
Author(s):  
Kozo Ishizaki ◽  
Kazuyuki Sawadaishi ◽  
Kazunobu Miura ◽  
Nariko Shinriki
Keyword(s):  
Escherichia Coli ◽  
Plasmid Dna

scholarly journals Cytoplasmic DNA can be detected by RNA fluorescence in situ hybridization

2019 ◽  
Vol 47 (18) ◽  
pp. e109-e109
Author(s):  
Eliraz Greenberg ◽  
Hodaya Hochberg-Laufer ◽  
Shalev Blanga ◽  
Noa Kinor ◽  
Yaron Shav-Tal
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dna Sequences ◽  
Plasmid Dna ◽  
Double Helix ◽  
Rna Molecules ◽  
Rna Fish ◽  
Dna Double Helix ◽  
Intracellular Detection

Abstract Fluorescence in situ hybridization (FISH) can be used for the intracellular detection of DNA or RNA molecules. The detection of DNA sequences by DNA FISH requires the denaturation of the DNA double helix to allow the hybridization of the fluorescent probe with DNA in a single stranded form. These hybridization conditions require high temperature and low pH that can damage RNA, and therefore RNA is not typically detectable by DNA FISH. In contrast, RNA FISH does not require a denaturation step since RNA is single stranded, and therefore DNA molecules are not detectable by RNA FISH. Hence, DNA FISH and RNA FISH are mutually exclusive. In this study, we show that plasmid DNA transiently transfected into cells is readily detectable in the cytoplasm by RNA FISH without need for denaturation, shortly after transfection and for several hours. The plasmids, however, are usually not detectable in the nucleus except when the plasmids are efficiently directed into the nucleus, which may imply a more open packaging state for DNA after transfection. This detection of plasmid DNA in the cytoplasm has implications for RNA FISH experiments and opens a window to study conditions when DNA is present in the cytoplasm.

Expression of recombinant DNA introduced into Chlamydia trachomatis by electroporation

1994 ◽  
Vol 40 (7) ◽  
pp. 583-591 ◽  
Author(s):  
Jeffrey E. Tam ◽  
Carolyn H. Davis ◽  
Priscilla B. Wyrick
Keyword(s):  
Chlamydia Trachomatis ◽  
Plasmid Dna ◽  
Southern Hybridization ◽  
Recombinant Dna ◽  
Chloramphenicol Acetyltransferase ◽  
Developmentally Regulated ◽  
Reticulate Body ◽  
Body Development ◽  
Coli Plasmid

Electroporation was used to introduce DNA into the elementary bodies of the obligate parasitic bacterium Chlamydia trachomatis. The source of DNA for these experiments was the chimeric plasmid pPBW100, which was constructed from the well-characterized 7.5-kb plasmid of C. trachomatis and the Escherichia coli plasmid pBGS9. To select directly for C. trachomatis carrying pPBW100, an in-frame gene fusion between the chlamydial promoter P7248 and a promoterless chloramphenicol acetyltransferase (cat) cassette was incorporated into the plasmid. After infection of McCoy cells with electroporated elementary bodies containing pPBW100, the following were observed: (i) the plasmid DNA was detected inside the chloramphenicol-resistant chlamydial inclusions by in situ and Southern hybridization analyses; (ii) both physical and biochemical evidence showed that chloramphenicol acetyltransferase was synthesized by the electroporated C. trachomatis; (iii) expression of P7248::cat was developmentally regulated and occurred during the early stages of chlamydial reticulate body development; and (iv) although the expression from P7248::cat was mainly transient, there were rare instances where chloramphenicol-resistant C. trachomatis were observed after four passages.Key words: chlamydia, electroporation, chimeric plasmid, expression.

Adsorption of Plasmid DNA ontoN,N‘- (Dimethylamino)ethyl-methacrylate Graft-Polymerized Poly-l-lactic Acid Film Surface for Promotion of in-Situ Gene Delivery

2007 ◽  
Vol 8 (6) ◽  
pp. 1951-1957 ◽  
Author(s):  
Tingting Jiang ◽  
Chang ◽  
Chunming Wang ◽  
Zhi Ding ◽  
Jiangning Chen ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Gene Delivery ◽  
Plasmid Dna ◽  
Film Surface ◽  
Ethyl Methacrylate

scholarly journals Exerting DNA Damaging Effects of the Ilimaquinones through the Active Hydroquinone Species

2021 ◽  
Vol 89 (2) ◽  
pp. 26
Author(s):  
Apisada Jiso ◽  
Laphatrada Yurasakpong ◽  
Sirorat Janta ◽  
Kulathida Chaithirayanon ◽  
Anuchit Plubrukarn
Keyword(s):  
Dna Damage ◽  
Plasmid Dna ◽  
Redox Cycling ◽  
Active Species ◽  
In Situ Reduction ◽  
Redox Transformation ◽  
Active Product ◽  
Cellular Dna ◽  
Damaging Effects

Possessing the quinone moiety, ilimaquinone (1), a sponge–derived sesquiterpene quinone, has been hypothesised to express its cytotoxicity through a redox cycling process, yielding active product(s) that can cause DNA damage. To determine the DNA damaging effects of 1 and examine whether a redox transformation may participate in its functions, the DNA damaging properties of 1, the corresponding hydroquinone (2) and hydroquinone triacetates (3) and their 5-epimeric counterparts (4–6) were tested and compared. When incubated directly with plasmid DNA, the hydroquinones were the only active species capable of cleaving the DNA. In cell-based assays, however, the quinones and hydroquinone triacetates were active in the same range as that of the corresponding hydroquinones, and all damaged the cellular DNA in a similar manner. The in situ reduction of 1 and 4 were supported by the decreases in the cytotoxicity when cells were pre-exposed to dicoumarol, an NAD(P)H:quinone oxidoreductase 1 (NQO1) inhibitor. The results confirmed the DNA damaging activities of the ilimaquinones 1 and 4, and indicated the necessity to undergo an in-situ transformation into the active hydroquinones, thereby exerting the DNA damaging properties as parts of the cytotoxic mechanisms.

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