Plasmid isolation

Tennstedt T, Szczepanowski R, Braun S et al (2003) Occurrence of integron-associated resistance gene cassettes located on antibiotic resistance plasmids isolated from a waste water treatment plant. EEMS Microbiol Ecol 45(3) 239-252... 

Schluter A, Szczepanowsld R, Puhler A et al (2007) Genomics of IncP-1 antibiotic resistance plasmids isolated from waste water treatment plants provides evidence for a widely accessible drug resistance gene pool. KEMS Microbiol Rev 31(4) 449-477... 

The instructor or teaching assistant can complete part A and provide cell cultures to students for plasmid isolation and electrophoresis. Approximately 3-4 hours are required for parts B and C. 

There are many published variations of the two most popular approaches to rapid plasmid isolation using either alkaline lysis43 or boiled... 

The plasmids isolated from clones No.6 and No. 16 were used to transform E.coli BL21 and W3110 strains. The cell cultivation conditions were similar to those applied for E.coli DH5 a cells. The accumulation of protein with molecular mass 72 kDa was demonstrated... 

A particularly useful laboratory application of restriction enzymes is in the epidemiologic investigation of bacterial and plasmid isolates to see if patient isolates reflect personnel- or equipment-based sources of infection (43). A fruitful approach to this issue has been to use restriction enzymes and probes to study the fingerprint pattern of ribosomal rRNA genes, which because of their Icirge number will demonstrate characteristic identifying patterns even within strains. 

JETSTAR Plasmid Kit (GENOMED) plasmid isolation kit, based alkaline lysis of the bacterial cell. Chromosomal DNA sticks to the cell wall and coprecipitates with the cell debris, whereas plasmid DNA remains solubilized. An anion exchange resin, provided in columns, binds the plasmid DNA under high salt conditions, the DNA is washed and eluted in low salt conditions. The solutions are provided in the kit El (cell resuspension buffer 50 mM Tris-HCl, pH 8.0, 10 mMEDTA), E2 (cell lysis solution 200 mM NaOH, 1% SDS), E3 (neutralization buffer 3.2 M KOAc/HOAc, pH 5.5), E4 (column equilibration... 

Dong, X.Q., Lindler, L.E., Chu, M.C. 2000. Complete DNA sequence and analysis of an emerging cryptic plasmid isolated from Yersinia pestis. Plasmid 43 144-148. 

Quantification of Plasmid Isolated from Transfected Cells... 

Delivery of the plasmid to the target cells is only useful if the plasmid remains intact during the process and it is in a transcription-ready form after cellular internalization. As indicated earlier, it is not clear yet which plasmid conformation(s) can be defined as the active species. Any analysis of plasmids isolated from cells or tissues should take this into consideration bearing... 

Plasmid isolation, restriction, cloning DNA sequencing, etc. were carried out, using standard protocols described by Sambrook et al. [22],... 

The degradative property of the strain BYAl was found to reside on a mega plasmid carried by it, which was confirmed by transforming an E. coli strain DHSa with the plasmid isolated fi om BYAl. The transformed E. coli could utilize BOAA as sole source of carbon and nitrogen, whereas the non-transformed cells could not. An overlapping restriction map of this plasmid, with respect to various enzymes, like HindUL, Kpnl, Pstl, and Notl [24], has been prepared (Figure 1),... 

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