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11.
David J Lee Lewis EH Bingle Karin Heurlier Mark J Pallen Charles W Penn Stephen JW Busby Jon L Hobman 《BMC microbiology》2009,9(1):252
Background
Homologous recombination mediated by the λ-Red genes is a common method for making chromosomal modifications in Escherichia coli. Several protocols have been developed that differ in the mechanisms by which DNA, carrying regions homologous to the chromosome, are delivered into the cell. A common technique is to electroporate linear DNA fragments into cells. Alternatively, DNA fragments are generated in vivo by digestion of a donor plasmid with a nuclease that does not cleave the host genome. In both cases the λ-Red gene products recombine homologous regions carried on the linear DNA fragments with the chromosome. We have successfully used both techniques to generate chromosomal mutations in E. coli K-12 strains. However, we have had limited success with these λ-Red based recombination techniques in pathogenic E. coli strains, which has led us to develop an enhanced protocol for recombineering in such strains. 相似文献12.
13.
JW Santo Domingo J C Radway E W Wilde P Hermann T C Hazen 《Journal of industrial microbiology & biotechnology》1997,18(6):389-395
Immobilization of the trichloroethylene-degrading bacterium Burkholderia cepacia was evaluated using hydrophilic polyurethane foam. The influence of several foam formulation parameters upon cell retention
was examined. Surfactant type was a major determinant of retention; a lecithin-based compound retained more cells than pluronic-
or silicone-based surfactants. Excessive amounts of surfactant led to increased washout of bacteria. Increasing the biomass
concentration in the foam from 4.8 to 10.5% dry weight per wet weight of foam resulted in fewer cells being washed out. Embedding
at reduced temperature did not significantly affect retention, while the use of a silane binding agent gave inconsistent
results. The optimal formulation retained all but 0.2% of total embedded cells during passage of 2 L of water through columns
containing 2 g of foam. All foam formulations tested reduced the culturability of embedded cells by several orders of magnitude,
but O2 consumption and CO2 evolution rates of embedded cells were never less than 50% of those of free cells. Nutrient amendments stimulated an increase
in cell volume and ribosomal activity in immobilized cells as indicated by hybridization studies using fluorescently labeled
ribosomal probes. These results indicate that, although immobilized cells were mostly nonculturable, they were metabolically
active and thus could be used for biodegradation of toxic compounds.
Received 23 December 1996/ Accepted in revised form 13 March 1997 相似文献
14.
Restriction-map variation associated with the G6PD polymorphism in natural populations of Drosophila melanogaster 总被引:10,自引:0,他引:10
Restriction-map variation was studied in 126 copies of the G6pd region in X
chromosome lines of Drosophila melanogaster from North America, Europe, and
Africa. Special attention was focused on the distribution of variation
relative to the geographically variable polymorphism for two
electrophoretic variants. Nucleotide heterozygosity as determined by eight
six-cutter restriction enzymes for the 13-kb region is estimated, on the
basis of the worldwide sample, to be 0.065%, which is the lowest value
reported for any comparable region in the D. melanogaster genome.
Significant linkage disequilibrium between electrophoretic alleles and
restriction-site variation is observed for several sites. In contrast to
published studies of other genetic regions, there are large insertions that
reach significant frequencies and are found across considerable geographic
distances. There is a clustering of this variation inside the first large
intervening sequence of the G6PD gene.
相似文献
15.
16.
Aartjan JW te Velthuis Jeroen F Admiraal Christoph P Bagowski 《BMC evolutionary biology》2007,7(1):129
Background
Development, differentiation and physiology of metazoans all depend on cell to cell communication and subsequent intracellular signal transduction. Often, these processes are orchestrated via sites of specialized cell-cell contact and involve receptors, adhesion molecules and scaffolding proteins. Several of these scaffolding proteins important for synaptic and cellular junctions belong to the large family of membrane-associated guanylate kinases (MAGUK). In order to elucidate the origin and the evolutionary history of the MAGUKs we investigated full-length cDNA, EST and genomic sequences of species in major phyla. 相似文献17.
18.
The growth and division of mitochondria during the cell cycle was investigated by a morphometric analysis of electron micrographs of synchronized HeLa cells. The ratio of total outer membrane contour length to cytoplasmic area did not vary significantly during the cell cycle, implying a continuous growth of the mitochondrial outer membrane. The mean fraction of cytoplasmic area occupied by mitochondrial profiles was likewise found to remain constant, indicating that the increase in total mitochondrial volume per cell occurs continuously during interphase, in such a way that the mitochondrial complement occupies a constant fraction( approximately 10-11(percent)) of the volume of the cytoplasm. The mean area, outer membrane contour length, and axis ratio of the mitochondrial profiles also did not vary appreciably during the cell cycle; furthermore, the close similarity of the frequency distributions of these parameters for the six experimental time-points suggested a stable mitochondrial shape distribution. The constancy of both the mean mitochondrial profile area and the number of mitochondrial profiles per unit of cytoplasmic area was interpreted to indicate the continuous division of mitochondria at the level of the cell population. Furthermore, no evidence was found for the occurrence of synchronous mitochondrial growth and division within individual cells. Thus, it appears that, in HeLa cells, there is no fixed temporal relationship between the growth and division of mitochondria and the events of the cell cycle. A number of statistical methods were developed for the purpose of making numerical estimates of certain three-dimensional cellular and mitochondrial parameters. Mean cellular and cytoplasmic volumes were calculated for the six time-points; both exhibited a nonlinear, approx. twofold increase. A comparison of the axis ratio distributions of the mitochondrial profiles with theoretical distributions expected from random sectioning of bodies of various three-dimensional shapes allowed the derivation of an "average" mitochondrial shape. This, in turn, permitted calculations to be made which expressed the two-dimensional results in three-dimensional terms. Thus, the estimated values for the number of mitochondria per unit of cytoplasmic volume and for the mean mitochondrial volume were found to remain constant during the cell cycle, while the estimated number of mitochondria per cell increase approx. twofold in an essentially continuous manner. 相似文献
19.
Premelting at the surface of ice crystals is caused by factors such as temperature, radius of curvature, and solute composition. When polycrystalline ice samples are warmed from well below the equilibrium melting point, surface melting may begin at temperatures as low as -15 degrees C. However, it has been reported (. Biophys. J. 65:1853-1865) that when polycrystalline ice was warmed in a differential scanning calorimetry (DSC) pan, melting began at about -50 degrees C, this extreme behavior being attributed to short-range forces. We show that there is no driving force for such premelting, and that for pure water samples in DSC pans curvature effects will cause premelting typically at just a few degrees below the equilibrium melting point. We also show that the rate of warming affects the slope of the DSC baseline and that this might be incorrectly interpreted as an endotherm. The work has consequences for DSC operators who use water as a standard in systems where subfreezing runs are important. 相似文献
20.
Pei-Li Yao Meng-Feng Tsai Yi-Chen Lin Chien-Hsun Wang Wei-Yu Liao Jeremy JW Chen Pan-Chyr Yang 《Respiratory research》2005,6(1):89