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排序方式: 共有74条查询结果,搜索用时 31 毫秒
1.
H Uhlmann V Beckert D Schwarz R Bernhardt 《Biochemical and biophysical research communications》1992,188(3):1131-1138
Expression systems for adrenodoxin into the periplasm and the cytoplasm of E. coli have been developed as a prerequisite for site-directed mutagenesis studies. In both systems the /2Fe-2S/ cluster of the protein was correctly assembled, the cytoplasmic one gives, however, a tenfold higher expression level. To determine which of the five cysteines at positions 46, 52, 55, 92, and 95 coordinate the /2Fe-2S/ center, they have been individually mutated into serines. From these mutants, only C95S forms a functionally active holoprotein. Thus, residues 46, 52, 55, and 92 are the cysteines that coordinate the /2Fe-2S/ cluster in adrenodoxin. 相似文献
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Summary The ability to recover male gametophyte derived plants, which is necessary to get transformed haploid plants, was verified for a hybrid of maize. Using the isolated microspore culture technique, a 9 × 10–5 plant regeneration frequency was obtained. Maize microspores were bombarded with tungsten particles using a PDS He/1000 apparatus. GUS expression in the microspores was maximum with 1.1 m diameter tungsten microprojectiles for 1100 and 1350 psi helium pressures at a 6 cm distance between the launch point and the target cells. Increasing the amount of DNA coated on the microparticles from 1.66 to 4 g DNA/mg of particles allowed a two-fold and four-fold increase of the GUS-expressing microspore frequency for 1100 and 1350 psi helium pressure bombardment, respectively. Optimal concentration of solidifying agent in the bombardment support culture medium was found to be 1%. Cell density ranging from 25000 microspores/bombardment to 100000 microspores/bombardment did not affect the frequency of GUS-expressing microspores. Using these optimal conditions, the maximum frequency of GUS-expressing microspores was found to be about 9 × 10–4, while maintaining an embryo formation frequency about 5 × 10–4.Abbreviations GUS
-glucuronidase
- PEG
polyethylene glycol 相似文献
5.
Genetic variation at the Major Histocompatibility Complex locus DQ beta was
analyzed in 233 beluga whales (Delphinapterus leucas) from seven
populations: St. Lawrence Estuary, eastern Beaufort Sea, eastern Chukchi
Sea, western Hudson Bay, eastern Hudson Bay, southeastern Baffin Island,
and High Arctic and in 12 narwhals (Monodon monoceros) sympatric with the
High Arctic beluga population. Variation was assessed by amplification of
the exon coding for the peptide binding region via the polymerase chain
reaction, followed by either cloning and DNA sequencing or single-stranded
conformation polymorphism analysis. Five alleles were found across the
beluga populations and one in the narwhal. Pairwise comparisons of these
alleles showed a 5:1 ratio of nonsynonymous to synonymous substitutions per
site leading to eight amino acid differences, five of which were
nonconservative substitutions, centered around positions previously shown
to be important for peptide binding. Although the amount of allelic
variation is low when compared with terrestrial mammals, the nature of the
substitutions in the peptide binding sites indicates an important role for
the DQ beta locus in the cellular immune response of beluga whales.
Comparisons of allele frequencies among populations show the High Arctic
population to be different (P < or = .005) from the other beluga
populations surveyed. In these other populations an allele, Dele-DQ
beta*0101-2, was found in 98% of the animals, while in the High Arctic it
was found in only 52% of the animals. Two other alleles were found at high
frequencies in the High Arctic population, one being very similar to the
single allele found in narwhal.
相似文献
6.
The slow-evolving mitochondrial DNAs of plants have potentially conserved information on the phylogenetic branching of the
earliest land plants. We present the nad2 gene structures in hornworts and liverworts and in the presumptive earliest-branching
vascular land plant clade, the Lycopodiopsida. Taken together with the recently obtained nad2 data for mosses, each class
of bryophytes presents another pattern of angiosperm-type introns conserved in nad2: intron nad2i1 in mosses; intron nad2i3
in liverworts; and both introns, nad2i3 and nad2i4, in hornworts. The lycopods Isoetes and Lycopodium show diverging intron
conservation and feature a unique novel intron, termed nad2i3b. Hence, mitochondrial introns in general are positionally stable
in the bryophytes and provide significant intraclade phylogenetic information, but the nad2 introns, in particular, cannot
resolve the interclade relationships of the bryophyte classes and to the tracheophytes. The necessity for RNA editing to reconstitute
conserved codon entities in nad2 is obvious for all clades except the marchantiid liverworts. Finally, we find that particularly
small group II introns appear as a general feature of the Isoetes chondriome. Plant mitochondrial peculiarities such as RNA
editing frequency, U-to-C type of RNA editing, and small group II introns appear to be genus-specific rather than gene-specific
features. 相似文献
7.
Analysis of lipocyte viability after liposuction 总被引:16,自引:0,他引:16
Boschert MT Beckert BW Puckett CL Concannon MJ 《Plastic and reconstructive surgery》2002,109(2):761-5; discussion 766-7
Free fat grafts from liposuction aspirate can be used as donor material for soft-tissue augmentation. The purpose of this study was to attempt to identify a subpopulation of adipose cells within liposuction aspirate with the greatest viability and, it is hoped, a greater chance for increased survival after transplantation. Liposuction samples were obtained from 20 individuals (16 women, four men; age range, 27 to 49 years). These samples were then centrifuged at 50 g. At 2-minute intervals, specimens from three different areas (superficial, middle, deep) were obtained from each specimen. After collagenase degradation, the specimens were stained with trypan blue, and the number of viable cells were counted. The bottom (deepest) layer consistently contained the highest number of viable cells after centrifugation: 250 percent more viable cells when compared with the top layer (p < 0.0001) and 140 percent more viable cells when compared with the middle layer (p < 0.0002). Centrifugation beyond 2 minutes did not increase the number or proportion of viable adipocytes. When using aspirated fat from liposuction for soft-tissue augmentation, centrifugation for 2 minutes at 50 g will stratify the adipocytes, with more viable cells being found at the deepest layer. Using only this bottom portion of the fat layer for transplantation will yield a fat graft with a greater number of viable adipocytes, potentially improving fat graft survival and decreased fat graft resorption. 相似文献
8.
Adamec J Beckert R Weiss D Klimesová V Waisser K Möllmann U Kaustová J Buchta V 《Bioorganic & medicinal chemistry》2007,15(8):2898-2906
New hybrid molecules of estrone were synthesized as compounds indicating promising biological activity (antibacterial, antimycobacterial, antifungal, and antiproliferative). The prepared molecules contained various heterocyclic units (pyridine, benzylsulfanyl derivatives of pyridine or derivatives of tetrazole) linked to estrone by n-heptyl bridges. The compounds with charge on molecule (the hybrid pyridinium or benzylsulfanylpyridinium salts) exhibited significant biological activity (antibacterial, antimycobacterial, antifungal, and antiproliferative). On the other hand, the compounds not in the form of salts (omega-(1-phenyl-5-tetrazolylthio)heptylethers of estrone) were inactive. The antimycobacterial activities of three different series of tetrazole derivatives (i.e., the hybrid molecules with estrone, tetrazole-5-thiols, and 5-benzylsulfanyl-1-phenyltetrazoles) with the same substituents on phenyl ring were compared. Amongst them, the 5-benzylsulfanyl-1-phenyltetrazoles were the most potent. 相似文献
9.
Sanja Cicko Melanie Grimm Korcan Ayata Jessica Beckert Anja Meyer Madelon Hossfeld Gernot Zissel Marco Idzko Tobias Müller 《Respiratory research》2015,16(1)
Rationale
Pulmonary fibrosis is a progressive disease with only few treatment options available at the moment. Recently, the nucleoside uridine has been shown to exert anti-inflammatory effects in different animal models, e.g. in acute lung injury or bronchial asthma.Method
Therefore, we investigated the influence of uridine supplementation on inflammation and fibrosis in the classical bleomycin model. Male C57BL/6 mice received an intratracheal injection of bleomycin on day 0 and were treated intraperitoneally with uridine or vehicle. The degree of inflammation and fibrosis was assessed at different time points.Results
Uridine administration resulted in attenuated inflammation, as demonstrated by reduced leukocytes and pro-inflammatory cytokines in the broncho-alveolar lavage (BAL) fluid. Furthermore, collagen deposition in the lung interstitium was also reduced by uridine supplementation. Similar results were obtained in a model in which animals received repeated intraperitoneal bleomycin injections. In addition uridine inhibited collagen and TGF-ß synthesis by primary lung fibroblasts, the release of pro-inflammatory cytokines by human lung epithelial cells, as well as the production of reactive oxygen species by human neutrophils.Conclusion
In summary, we were able to show that uridine has potent anti-inflammatory and anti-fibrotic properties. As uridine supplementation has been shown to be well tolerated and safe in humans, this might be a new therapeutic approach for the treatment of fibrotic lung diseases. 相似文献10.
Enhancement of maize transformation efficiency by the use of maize matrix attachment regions 总被引:2,自引:0,他引:2
Fran?ois?TorneyEmail author Anne?Partier Pierre?Barret Michel?Beckert 《Plant Cell, Tissue and Organ Culture》2005,80(3):295-303
Improving genetic transformation efficiency is a major concern in plant genetic engineering. While various strategies have been investigated, the enhancement of selectable marker gene expression has not been tried extensively. We used maize matrix attachment regions (MARs) to bracket an herbicide resistance transgene, bar. MARs have been reported to enhance transgene expression level and stability. We show here that MARs not only enhance transformation efficiency by 50%, but are also able to increase or decrease relative efficiencies of each step of the regeneration process depending on MAR sequence combinations. Furthermore, we assessed the trans-effect of MARs in co-bombardment experiments with two independent plasmids, one including the MAR sequences and the other one the bar gene. As for simple bombardment, MARs enhanced transformation efficiency by having a positive influence on organogenesis step in the regeneration process. 相似文献