Chromosomal DNA from Coxiella burnetii Scurry Q217 was screened for the presence of plasmid-homologous sequences. Total DNA from Scurry Q217 was digested with NotI, and the resulting DNA fragments were separated by contour-clamped homogeneous electric field pulsed-field gel electrophoresis (CHEF-PFGE). Following hybridization with biotin-labeled QpH1 plasmid as a probe, two DNA fragments of 40 and 170 kb were identified as targets. These fragments were cloned, and subclones containing QpH1-homologous sequences were completely sequenced. The physical mapping of DNA fragments was achieved by PCR with primers derived from adjacent fragments, and a total of 18,360 bp was sequenced. ORFs were different in size, 6 ORFs were newly generated, and 25 ORFs were lost. It was found that plasmid-homologous sequences in Scurry Q217 were of chromosomal origin.Coxiella burnetii, the only member of the genus Coxiella, is the causative agent of Q fever in humans as well as of abortions in domestic animals, especially ruminants. So far, different criteria have been applied for the classification of C. burnetii isolates. Samuel et al. (7) distinguish between so-called acute and chronic isolates that cause acute or chronic Q fever in humans, apparently depending on the plasmid type. However, recent experimental findings (3) and PCR analysis of C. burnetii strains isolated from patients exhibiting chronic Q fever (11) revealed that isolates containing either QpH1 or QpRS can cause endocarditis. Yu and Raoult (15) confirmed these results by serotyping isolates with monoclonal antibodies and postulated that the development of acute or chronic Q fever depends on the patient's condition and immune status rather than on the plasmid type of the causative agent. Nevertheless, plasmids are still considered to be of major importance concerning virulence factors, mechanisms of pathogenicity, and proliferation in the phagolysosomal compartment.Assuming that the plasmid-coded sequences are of essential importance for C. burnetii, the existence of the plasmidless C. burnetii isolate Scurry Q217 (8) was rather unexpected. However, it was shown that Scurry Q217 contains chromosomally integrated plasmid-homologous DNA sequences (8). In an attempt to understand the significance of plasmids in C. burnetii, we examined plasmidless isolates with chromosomally integrated plasmid-homologous DNA fragments. MATERIALS AND METHODSDNA-modifying enzymes. Restriction endonucleases and other DNA-modifying enzymes were purchased from Stratagene (Heidelberg, Germany) and Amersham (Bad Homburg, Germany) and used under conditions recommended by the manufacturers.Plasmids and bacterial strains. C. burnetii DNA fragments were cloned in phagemid vector pBluescript II KS(ϩ), cosmid vector SuperCos 1, or lambda phage vector EMBL4 (Stratagene). Recombinant cosmid and lambda phage DNAs were packaged (Gigapack II packaging extract; Stratagene) and transfected into Escherichia coli XL1 Blue (Stratagene). C. burnetii reference isolates Scurry Q217, Priscilla Q177, Dugway 7E22-...

Copper and superoxide are used by the phagocytes to kill bacteria. Copper is a host effector encountered by uropathogenic Escherichia coli (UPEC) during urinary tract infection in a non-human primate model, and in humans. UPEC is exposed to higher levels of copper in the gut prior to entering the urinary tract. Effects of pre-exposure to copper on bacterial killing by superoxide has not been reported. We hypothesized that copper-replete E. coli is more sensitive to killing by superoxide in vitro, and in activated macrophages. We utilized wild-type UPEC strain CFT073, and its isogenic mutants lacking copper efflux systems, superoxide dismutases (SODs), regulators of a superoxide dismutase, and complemented mutants to address this question. Surprisingly, our results reveal that copper protects UPEC against killing by superoxide in vitro. This copper-dependent protection was amplified in the mutants lacking copper efflux systems. Increased levels of copper and manganese were detected in UPEC exposed to sublethal concentration of copper. Copper activated the transcription of sodA in a SoxR-and SoxS-dependent manner resulting in enhanced levels of SodA activity. Importantly, pre-exposure to copper increased the survival of UPEC within RAW264.7 and bone marrow-derived murine macrophages. Loss of SodA, but not SodB or SodC, in UPEC obliterated copper-dependent protection from superoxide in vitro, and from killing within macrophages. Collectively, our results suggest a model in which sublethal levels of copper trigger the activation of SodA and SodC through independent mechanisms that converge to promote the survival of UPEC from killing by superoxide. A major implication of our findings is that bacteria colonizing copper-rich milieus are primed for efficient detoxification of superoxide.

Siphophage Staley was isolated because of its ability to grow on Bacillus megaterium. Here we report the complete genome and annotation of phage Staley and describe core features. Among its interesting genes is one encoding an SleB germination protein.

The complete annotated genome sequence of Bacillus megaterium bacteriophage Slash is described here. Several key features related to morphogenesis, replication/recombination, DNA metabolism, and lysis are described. Slash also encodes a homolog of SleB, a germination-specific cell wall amidase.

I. INTRODUCTION peneal (pn'al) adj. [[< Gr poini, penalty]] of, for, constituting, or deserving punishment 1 penecil (pen'sal) n. [[< L penis, tail]] a rod-shaped instrument with a core of graphite, crayon, etc. that is sharpened to a point for writing, etc. 2 peenis (pnis) n .... [[L]] the male organ of sexual intercourse 3 My fingers rest uneasily upon the keyboard of my electronic pen as I reflect upon how to write a paper regarding Women and Law. 4 I am sensitive to being male-a white, heterosexual malethoroughly steeped (both culturally and academically) in the legal, philosophical, and theological heritage of the West. 5 Feminist thought has taught me that all things said are said from a particular perspective. 6 More profoundly, feminist thought has taught me that gender identity may well inform the very structure of thought from which one's perspective is articulated. 7 In large measure, this