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41.
Shibin Gao Carlos Martinez Debra J. Skinner Alan F. Krivanek Jonathan H. Crouch Yunbi Xu 《Molecular breeding : new strategies in plant improvement》2008,22(3):477-494
Leaf collection from the field, labeling and tracking back to the source plants after genotyping are rate limiting steps in
leaf DNA-based genotyping. In this study, an optimized genotyping method using endosperm DNA sampled from single maize seeds
was developed, which can be used to replace leaf DNA-based genotyping for both genetic studies and breeding applications.
A similar approach is likely to be suitable for all plants with relatively large seeds. Part of the endosperm was excised
from imbibed maize seeds and DNA extracted in 96-tube plates using individuals from eight F2 populations and seven inbreds. The quality of the resultant DNA was functionally comparable to DNA extracted from leaf tissue.
Extraction from 30 mg of endosperm yields 3–10 μg DNA, which is sufficient for analysis of 200–400 agarose-gel PCR-based markers,
with the potential for several million chip-based SNP marker analyses. By comparing endosperm DNA and leaf DNA for individuals
from an F2 population, genotyping errors caused by pericarp contamination and hetero-fertilization were found to average 3.8 and 0.6%,
respectively. Endosperm sampling did not affect germination rates under controlled conditions, although under normal field
conditions the germination rate, seedling establishment, and growth vigor were significantly lower than that of non-sampled
controls for some genotypes. However, careful field management can compensate for these effects. Seed DNA-based genotyping
lowered costs by 24.6% compared to leaf DNA-based genotyping due to reduced field plantings and labor costs. A substantial
advantage of this approach is that it can be used to select desirable genotypes before planting. As such it provides an opportunity
for dramatic improvements in the efficiency and selective gain of breeding systems based on optimum combinations of marker-assisted
selection and phenotypic selection within and between generations. 相似文献
42.
The effects of K+ concentration, light intensity and CO2 levels on the volume of Commelina communis L. guard cell protoplasts were studied. Two degrees of swelling response were observed, both dependent on an external supply of K+ , but not necessarily on the supply of a permeant anion. The presence of K+ itself, independent of light or CO2 level, stimulated swelling at a relatively slow rate. When K+ , light and low CO2 conditions were supplied together, the swelling was relatively rapid and of high magnitude. The rapid swelling was specific for K+ and Rb+ giving a half maximal effect after 2 h at a KCl concentration of about 18 mmol m−3 . The addition of CaCl2 at 1 mol m−3 inhibited K+ -dependent swelling under all conditions tested. The response to light and low CO2 levels by Commelina guard cell protoplasts is thought to reflect a high degree of physiological integrity. 相似文献
43.
Enzyme activation for organic solvents made easy 总被引:1,自引:0,他引:1
Enzymes are highly selective catalysts that perform intricate chemistries at ambient temperatures and pressures. Although water is the solvent of life, it is a poor solvent for most synthetic organic reactions and, therefore, most chemists avoid aqueous solutions for synthetic applications. However, when removed from the aqueous environment and placed in an organic solvent, enzyme activity is reduced greatly. Here, we present a general overview of recent efforts to activate enzymes for use in nonaqueous media, giving particular focus to the use of simple salts as additives that result in significant biocatalytic improvements. 相似文献
44.
ED Dean LM Mexas NL Cápiro JE McKeon MR DeLong KD Pennell JA Doorn V Tangpricha GW Miller ML Evatt 《PloS one》2012,7(7):e39227
Recent clinical evidence supports a link between 25-hydroxyvitamin D insufficiency (serum 25-hydroxyvitamin D [25(OH)D] levels <30 ng/mL) and Parkinson's disease. To investigate the effect of 25(OH)D depletion on neuronal susceptibility to toxic insult, we induced a state of 25(OH)D deficiency in mice and then challenged them with the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). We found there was no significant difference between control and 25(OH)D-deficient animals in striatal dopamine levels or dopamine transporter and tyrosine hydroxylase expression after lesioning with MPTP. Additionally, we found no difference in tyrosine hydroxylase expression in the substantia nigra pars compacta. Our data suggest that reducing 25(OH)D serum levels in mice has no effect on the vulnerability of nigral dopaminergic neurons in vivo in this model system of parkinsonism. 相似文献
45.
Eftimie R Dushoff J Bridle BW Bramson JL Earn DJ 《Bulletin of mathematical biology》2011,73(12):2932-2961
Recent advances in virology, gene therapy, and molecular and cell biology have provided insight into the mechanisms through
which viruses can boost the anti-tumor immune response, or can infect and directly kill tumor cells. A recent experimental
report (Bridle et al. in Molec. Ther. 18(8):1430–1439, 2010) showed that a sequential treatment approach that involves two viruses that carry the same tumor antigen leads to an improved
anti-tumor response compared to the effect of each virus alone. In this article, we derive a mathematical model to investigate
the anti-tumor effect of two viruses, and their interactions with the immune cells. We discuss the conditions necessary for
permanent tumor elimination and, in this context, we stress the importance of investigating the long-term effect of non-linear
interactions. In particular, we discuss multi-stability and multi-instability, two complex phenomena that can cause abrupt
transitions between different states in biological and physical systems. In the context of cancer immunotherapies, the transitions
between a tumor-free and a tumor-present state have so far been associated with the multi-stability phenomenon. Here, we show
that multi-instability can also cause the system to switch from one state to the other. In addition, we show that the multi-stability
is driven by the immune response, while the multi-instability is driven by the presence of the virus. 相似文献
46.
Jonathan W. Yewdell 《Molecular & cellular proteomics : MCP》2022,21(7):100230
In the 35 years since the revelation that short peptides bound to major histocompatibility complex class I and II molecules are the secret of the major histocompatibility complex–restricted nature of T-cell recognition, there has been enormous progress in characterizing the immunopeptidome, the repertoire of peptide presented for immunosurveillance. Here, the major milestones in the journey are marked, the contribution of proteasome-mediated splicing to the immunopeptidome is discussed, and exciting recent findings relating the immunopeptidome to the translatome revealed by ribosome profiling (RiboSeq) is detailed. Finally, what is needed for continued progress is opined about, which includes the infusion of talented young scientists into the antigen-processing field, currently undergoing a renaissance; thanks in part to the astounding success of T-cell–based cancer immunotherapy. 相似文献
47.
48.
49.
Holleran JP Glover ML Peters KW Bertrand CA Watkins SC Jarvik JW Frizzell RA 《Molecular medicine (Cambridge, Mass.)》2012,18(1):685-696
Numerous human diseases arise because of defects in protein folding, leading to their degradation in the endoplasmic reticulum. Among them is cystic fibrosis (CF), caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR ), an epithelial anion channel. The most common mutation, F508del, disrupts CFTR folding, which blocks its trafficking to the plasma membrane. We developed a fluorescence detection platform using fluorogen-activating proteins (FAPs) to directly detect FAP-CFTR trafficking to the cell surface using a cell-impermeant probe. By using this approach, we determined the efficacy of new corrector compounds, both alone and in combination, to rescue F508del-CFTR to the plasma membrane. Combinations of correctors produced additive or synergistic effects, improving the density of mutant CFTR at the cell surface up to ninefold over a single-compound treatment. The results correlated closely with assays of stimulated anion transport performed in polarized human bronchial epithelia that endogenously express F508del-CFTR. These findings indicate that the FAP-tagged constructs faithfully report mutant CFTR correction activity and that this approach should be useful as a screening assay in diseases that impair protein trafficking to the cell surface. 相似文献
50.
Risks and benefit evaluation for controlled human infection studies, where healthy volunteers are deliberately exposed to infectious agents to evaluate vaccine efficacy, should be explicit, systematic, thorough, and non-arbitrary. Decision analysis promotes these qualities using four steps: (1) determining explicit criteria and measures for evaluation, (2) identifying alternatives to the study, (3) defining the models used to estimate the measures for each alternative, and (4) running the models to produce the estimates and compare the alternatives. In this paper, we describe how decision analysis might be applied by funders and regulators, as well as by others contemplating the use of novel controlled human infection studies for vaccine development and evaluation. 相似文献