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排序方式: 共有223条查询结果,搜索用时 31 毫秒
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93.
P R Hawkins M T Runnegar A R Jackson I R Falconer 《Applied and environmental microbiology》1985,50(5):1292-1295
Cylindrospermopsis raciborskii, a tropical blooming species of cyanobacterium (blue-green alga), was isolated from the domestic water supply reservoir on Palm Island, a continental island off the tropical northeast coast of Australia. This species, not previously known to be toxic, was shown to be severely hepatotoxic for mice. The 50% lethal dose at 24 h after injection was found to be 64 +/- 5 mg of freeze-dried culture per kg of mouse. The principal lesion produced was centrilobular to massive hepatocyte necrosis, but various degrees of injury were also seen in the kidneys, adrenal glands, lungs, and intestine. The possible implication of this finding in relation to an incident of hepatoenteritis in humans living on the island is discussed. 相似文献
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95.
Kaar W Hartmann BM Fan Y Zeng B Lua LH Dexter AF Falconer RJ Middelberg AP 《Biotechnology and bioengineering》2009,102(1):176-187
Biosurfactants have been the subject of recent interest as sustainable alternatives to petroleum-derived compounds in areas ranging from soil remediation to personal and health care. The production of naturally occurring biosurfactants depends on the presence of complex feed sources during microbial growth and requires multicomponent enzymes for synthesis within the cells. Conversely, designed peptide surfactants can be produced recombinantly in microbial systems, enabling the generation of improved variants by simple genetic manipulation. However, inefficient downstream processing is still an obstacle for the biological production of small peptides. We present the production of the peptide biosurfactant GAM1 in recombinant E. coli. Expression was performed in fusion to maltose binding protein using chemically defined minimal medium, followed by a single-step affinity capture and enzymatic cleavage using tobacco etch virus protease. Different approaches to the isolation of peptide after cleavage were investigated, with special emphasis on rapid and simple procedures. Solvent-, acid-, and heat-mediated precipitation of impurities were successfully applied as alternatives to post-cleavage chromatographic peptide purification, and gave peptide purities exceeding 90%. Acid precipitation was the method of choice, due to its simplicity and the high purification factor and recovery rate achieved here. The functionality of the bio-produced peptide was tested to ensure that the resulting peptide biosurfactant was both surface active and able to be triggered to switch between foam-stabilizing and foam-destabilizing states. 相似文献
96.
Lisa M. Seymour Ania T. Deutscher Cheryl Jenkins Tracey A. Kuit Linda Falconer F. Chris Minion Ben Crossett Matthew Padula Nicholas E. Dixon Steven P. Djordjevic Mark J. Walker 《The Journal of biological chemistry》2010,285(44):33971-33978
Porcine enzootic pneumonia is a chronic respiratory disease that affects swine. The etiological agent of the disease, Mycoplasma hyopneumoniae, is a bacterium that adheres to cilia of the swine respiratory tract, resulting in loss of cilia and epithelial cell damage. A M. hyopneumoniae protein P116, encoded by mhp108, was investigated as a potential adhesin. Examination of P116 expression using proteomic analyses observed P116 as a full-length protein and also as fragments, ranging from 17 to 70 kDa in size. A variety of pathogenic bacterial species have been shown to bind the extracellular matrix component fibronectin as an adherence mechanism. M. hyopneumoniae cells were found to bind fibronectin in a dose-dependent and saturable manner. Surface plasmon resonance was used to show that a recombinant C-terminal domain of P116 bound fibronectin at physiologically relevant concentrations (KD 24 ± 6 nm). Plasmin(ogen)-binding proteins are also expressed by many bacterial pathogens, facilitating extracellular matrix degradation. M. hyopneumoniae cells were found to also bind plasminogen in a dose-dependent and saturable manner; the C-terminal domain of P116 binds to plasminogen (KD 44 ± 5 nm). Plasminogen binding was abolished when the C-terminal lysine of P116 was deleted, implicating this residue as part of the plasminogen binding site. P116 fragments adhere to the PK15 porcine kidney epithelial-like cell line and swine respiratory cilia. Collectively these data suggest that P116 is an important adhesin and virulence factor of M. hyopneumoniae. 相似文献
97.
Parker J. Nichols Isaac Falconer Aaron Griffin Colin Mant Robert Hodges Christopher J. McKnight Beat Vgeli Liliya Vugmeyster 《Protein science : a publication of the Protein Society》2020,29(7):1641-1654
We have investigated the effect of deuteration of non‐exchangeable protons on protein global thermal stability, hydrophobicity, and local flexibility using well‐known thermostable model systems such as the villin headpiece subdomain (HP36) and the third immunoglobulin G‐binding domain of protein G (GB3). Reversed‐phase high‐performance liquid chromatography (RP‐HPLC) measurements as a function of temperature probe global thermal stability in the presence of acetonitrile, while differential scanning calorimetry determines thermal stability in solution. Both indicate small but measurable changes in the order of several degrees. RP‐HPLC also permitted quantification of the effect of deuteration of just three core phenylalanine side chains of HP36. NMR dynamics investigation has focused on methyl axes motions using cross‐correlated relaxation measurements. The analysis of order parameters provided a complex picture indicating that deuteration generally increases motional amplitudes of sub‐nanosecond motion in GB3 but decreases those in HP36. Combined with earlier dynamics measurements at Cα–Cβ sites and backbone sites of GB3, which probed slower time scales, the results point to the need to probe multiple atoms in the protein and variety of time scales to the discern the full complexity of the effects of deuteration on dynamics. 相似文献
98.
D Purnomosari T Aryandono K Setiaji SB Nugraha G Pals 《Biotechnic & histochemistry》2013,88(2-3):79-85
The HER-2/neu transmembrane tyrosine kinase receptor is both a prognostic marker and a therapeutic target for breast cancer. Accurate determination of HER-2/neu status is a prerequisite for selecting breast tumors for HER-2/neu immunotherapy or for taxan based chemotherapy. Unfortunately, there is no consensus concerning how this determination should be reached. We compared assessment of HER-2/neu status using Multiplex ligation-dependent probe amplification (MLPA) and immunohistochemistry (IHC). The patient group comprised 60 Indonesian breast cancers patients. IHC was performed on paraffin sections using the CB11 antibody from Novocastra. Results were scored according to the Hercept test. For MLPA, DNA was extracted from frozen samples, PCR amplified with a probe set containing three hemi-primer sets for the HER-2 locus and another nine control probes spread over chromosome 17 and other chromosomes, and analyzed on a gene scanner. A ratio above two for at least two HER-2 locus probes compared to the control probes was regarded as amplification. IHC for HER-2/neu was negative in 36 cases, and 24 cases (40%) showed expression. Seven, eight and nine of the latter cases were 1+, 2+ and 3+ positive, respectively. Forty-seven cases showed no amplification by MLPA, and 13 cases (22%) were amplified. Comparison of IHC and MPLA showed that none of the 36 IHC-negative or seven IHC 1+ cases was amplified. Five of the eight (63%) 2+ cases were amplified, and eight of nine (89%) of the IHC 3+ tumors showed gene amplification by MLPA assay. For HER-2/neu, there is a good correlation between gene amplification detected by MLPA and overexpression by IHC in invasive breast cancer. It appears that MLPA can detect the HER-2 amplified cases in the IHC 2+ class. Because MLPA is quick and inexpensive, it is an attractive method for detecting HER-2/neu amplification in daily laboratory practice. 相似文献
99.
Leuconostoc mesenteroides B-512FMC dextransucrase was found to synthesize dextrans of varying molecular weights by selecting the concentrations of dextransucrase and sucrose, as well as the temperature. Four enzyme concentrations (50, 10, 1.0, and 0.1 U/mL), five sucrose concentrations (20, 50, 100, 200 and 1000 mM), and two temperatures (20 °C and 30 °C) were studied. The highest amount of enzyme (50 U/mL), with the lowest concentration of sucrose (20 mM), and the lower temperature of 20 °C gave the lowest number-average molecular weight (MWn) of 20,630 Da, respectively. As the sucrose concentration was increased, 50 mM, 100 mM, and 200 mM, the MWn was 49,240 Da, 63,350 Da, and 126,720 Da, respectively. The next enzyme concentration (10 U/mL) gave a similar upward trend, starting at 73,130 Da and ending at 237,870 Da at 20 °C and 130,040 Da and ending at 415,770 Da at 30 °C. The upward trend continued for the 1.0 and 0.1 U/mL enzyme concentrations. An increase in the temperature had the overall effect of increasing the MWn for each decreasing concentration of enzyme and increasing concentration of sucrose. For 0.1 U/mL and 1000 mM sucrose at 30 °C, the MWn was 1,645,700 Da. The results of the study show that the molecular weights of the synthesized dextrans were inversely proportional to the concentration of the enzyme and directly proportional to the concentration of sucrose and the temperature. 相似文献
100.
Linda Irvine Donald W. Falconer Claire Jones Ian W. Ricketts Brian Williams Iain K. Crombie 《PloS one》2012,7(12)