Early in F plasmid conjugative transfer, the F relaxase, TraI, cleaves one plasmid strand at a site within the origin of transfer called nic. The reaction covalently links TraI Tyr16 to the 5'-ssDNA phosphate. Ultimately, TraI reverses the cleavage reaction to circularize the plasmid strand. The joining reaction requires a ssDNA 3'-hydroxyl; a second cleavage reaction at nic, regenerated by ext...

In combination with novel linear covalently closed (LCC) DNA minivectors, referred to as DNA ministrings, a gemini surfactant-based synthetic vector for gene delivery has been shown to exhibit enhanced delivery and bioavailability while offering a heightened safety profile. Due to topological differences from conventional circular covalently closed (CCC) plasmid DNA vectors, the linear topology...

The QuikChange™ site-directed mutagenesis method is popular but imperfect. An improvement by using partially overlapping primers has been reported several times; however, it is incompatible with the proposed mechanism. The QuikChange™ method using complementary primers is proposed to linearly amplify a target plasmid with the products annealing to produce double-stranded DNA molecules with 5'-o...

The majority of the human population harbors latent cytomegalovirus. Although CD141 peripheral blood mononuclear cells have been implicated as sites of latency, the conformation of the latent viral genome in these cells is unknown. In this study, the conformation of viral genomic DNA was assessed in CD141 cells from healthy virus seropositive carriers using an electrophoretic separation on nati...

Division of Molecular Immunology, and Neurobioiogy, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada MSG 1XS Here we describe a modificat ion of the polymerase chain reaction--junctionPCR IJ-PCR)--that has been designed to facilitate the identification and subsequent screening of transgenic animals and their offspring. This procedure targets a unique recombina...

We have studied the replication of plasmids composed of bovine papillomavirus type 1 (BPV1) origin of replication and expression cartridges for viral proteins E1 and E2 in hamster and mouse cells. We found that the replication mode changed dramatically at different expression levels of the E1 protein. At high levels of the E1 protein, overreplication of the origin region of the plasmid was obse...

Plasmid cp8.3 of Borrelia afzelii IP21 carries several open reading frames (ORFs) and a 184-bp inverted repeat (IR) element. It has been speculated that this plasmid may encode factors involved in virulence or infectivity. In this report, we have characterized the distribution, molecular variability, and organization of ORFs 1, 2, and 4 and the IR elements among isolates of the Borrelia burgdor...

The nucleotide sequence of Streptomyces lividans linear plasmid SLP2 consists of 50,410 bp (C. H. Huang, C. Y. Chen, H. H. Tsai, C. Chen, Y. S. Lin, and C. W. Chen, Mol. Microbiol. 47:1563-1576, 2003). Here we report that the basic SLP2 locus for plasmid replication in circular mode resembles that of Streptomyces linear plasmids pSLA2 and SCP1 and comprises iterons(SLP2) and the adjacent rep(SL...

Background: Stereotactic radiosurgery is an important clinical tool for the treatment of small lesions in the brain, including benign conditions, malignant and localized metastatic tumors. A dosimetry study was performed for Elekta ‘Synergy S ’ as a dedicated Stereotactic radiosurgery unit, capable of generating circular radiation fields with diameters of 1-5 cm at isocentre using the BEAM/EGS4...

side weir is a flow control structure that used extensively in irrigation and drainage as well as sewer networks. throughout the present research, a study comprised of 162 experiments was done on sharp-crested semi-circular side weir. since the height in this side weir varies along its length, this feature enables it to more properly control flood than rectangular side weirs in various flood co...