17 resultados para restriction sites
Resumo:
Campylobacter jejuni is a leading cause of human diarrheal illness in the world, and research on it has benefitted greatly by the completion of several genome sequences and the development of molecular biology tools. However, many hurdles remain for a full understanding of this unique bacterial pathogen. One of the most commonly used strains for genetic work with C. jejuni is NCTC11168. While this strain is readily transformable with DNA for genomic recombination, transformation with plasmids is problematic. In this study, we have identified a determinant of this to be cj1051c, predicted to encode a restriction-modification type IIG enzyme. Knockout mutagenesis of this gene resulted in a strain with a 1,000-fold-enhanced transformation efficiency with a plasmid purified from a C. jejuni host. Additionally, this mutation conferred the ability to be transformed by plasmids isolated from an Escherichia coli host. Sequence analysis suggested a high level of variability of the specificity domain between strains and that this gene may be subject to phase variation. We provide evidence that cj1051c is active in NCTC11168 and behaves as expected for a type IIG enzyme. The identification of this determinant provides a greater understanding of the molecular biology of C. jejuni as well as a tool for plasmid work with strain NCTC11168. © 2012, American Society for Microbiology.
Resumo:
Cement-bentonite (CB) cutoff walls have long been used to control ground water flow and contaminant migration at polluted sites. Hydraulic conductivity and unconfined compressive strength are two short-term properties often used by industry and owners in CB specification and are important parameters discussed in this paper. For polluted sites, long-term compatibility is also an important issue. These properties are coupled to a number of external factors including the mix design, construction sequence, presence/absence of contaminants at the site. Additional short-term properties for engineering assessment include the stressstrain characteristics in both drained and undrained shear in both with and without confinement as well as one-dimensional consolidation properties. Long term CB properties are affected by aging, reaction chemistry, drying, in situ stress state, and interaction with the polluted environment. © 2013 Taylor & Francis Group.