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41.
Signaling proteins from the same family can have markedly different roles in a given cellular context. Here, we show that expression of one hundred constitutively active human small GTPases induced cell morphologies that fell into nine distinct classes. We developed an algorithm for pairs of classes that predicted amino acid positions that can be exchanged to create mutants with switched functionality. The algorithm was validated by creating switch-of-function mutants for Rac1, CDC42, H-Ras, RalA, Rap2B, and R-Ras3. Contrary to expectations, the relevant residues were mostly outside known interaction surfaces and were structurally far apart from each other. Our study shows that specificity in protein families can be explored by combining genome-wide experimental functional classification with the creation of switch-of-function mutants. 相似文献
42.
Testing the robustness of the new Haseman-Elston quantitative-trait loci-mapping procedure 下载免费PDF全文
Variance components (VC) techniques have emerged as among the more powerful methods for detection of quantitative-trait loci (QTL) in linkage analysis. Allison et al. found that, with particularly marked leptokurtosis in the phenotypic distribution and moderate-to-high residual sibling correlation, maximum likelihood (ML) VC methods may produce a severe excess of type I errors. The new Haseman-Elston (NHE) method is a least-squares-based VC method for mapping of QTL in sib pairs (Elston et al.). Using simulation, we investigate the robustness of the NHE to marked nonnormality, by means of the same distributions and worst-case conditions identified by Allison et al. for the ML approach (i.e., 100 pairs; high residual sibling correlation). Results showed that, when marked nonnormality is present, the NHE can be used without severe type I error-rate inflation, even at very small alpha levels. 相似文献
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Phage libraries displaying cDNA or random peptides have been used for profiling autoantibodies in cancer. The detection of
autoantibodies in human sera using phages displaying specific epitopes is usually performed by phage-immobilized ELISAs which
can detect specific antibodies without identification of whole antigens. However, these ELISAs can give feeble detection signals
that are indistinguishable from background signals which are caused by human sera. To improve the usefulness of phage ELISA
for human sera, the conditions for each step in phage ELISA were optimized. The antigenicity of phage antigens was maximal
when using coating buffer of neutral pH. By using protein-free blocking buffer and pre-adsorbing human sera with phage host
cell ER2738 extracts significantly decreased non-specific signals. Finally, when these conditions were applied to phage ELISA
using K10P1, the values of the negative controls were concentrated near cutoff values, which made the assay more reliable.
The optimized phage ELISA conditions described here would increase the efficacy of detection specific autoantibodies in human
sera. 相似文献
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Embryonic germ (EG) cells are undifferentiated stem cells isolated from cultured primordial germ cells (PGC). Porcine EG cell lines with capacities of both in vitro and in vivo differentiation have been established. Because EG cells can be cultured indefinitely in an undifferentiated state, they may be more suitable for nuclear donor cells in nuclear transfer (NT) than somatic cells that have limited lifespan in primary culture. Use of EG cells could be particularly advantageous to provide an inexhaustible source of transgenic cells for NT. In this study the efficiencies of transgenesis and NT using porcine fetal fibroblasts and EG cells were compared. The rate of development to the blastocyst stage was significantly higher in EG cell NT than somatic cell NT (94 of 518, 18.2% vs. 72 of 501, 14.4%). To investigate if EG cells can be used for transgenesis in pigs, green fluorescent protein (GFP) gene was introduced into porcine EG cells. Nuclear transfer embryos using transfected EG cells gave rise to blastocysts (29 of 137, 21.2%) expressing GFP based on observation under fluorescence microscope. The results obtained from the present study suggest that EG cell NT may have advantages over somatic cell NT, and transgenic pigs may be produced using EG cells. 相似文献
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Benedetta Artegiani Lisa van Voorthuijsen Rik G.H. Lindeboom Daniëlle Seinstra Inha Heo Pablo Tapia Carmen López-Iglesias Daniel Postrach Talya Dayton Rurika Oka Huili Hu Ruben van Boxtel Johan H. van Es Johan Offerhaus Peter J. Peters Jacco van Rheenen Michiel Vermeulen Hans Clevers 《Cell Stem Cell》2019,24(6):927-943.e6
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Seung Yeon Shin Soochahn Lee Il Dong Yun Ho Yub Jung Yong Seok Heo Sun Mi Kim Kyoung Mu Lee 《PloS one》2015,10(12)
In this paper, we present a novel cascaded classification framework for automatic detection of individual and clusters of microcalcifications (μC). Our framework comprises three classification stages: i) a random forest (RF) classifier for simple features capturing the second order local structure of individual μCs, where non-μC pixels in the target mammogram are efficiently eliminated; ii) a more complex discriminative restricted Boltzmann machine (DRBM) classifier for μC candidates determined in the RF stage, which automatically learns the detailed morphology of μC appearances for improved discriminative power; and iii) a detector to detect clusters of μCs from the individual μC detection results, using two different criteria. From the two-stage RF-DRBM classifier, we are able to distinguish μCs using explicitly computed features, as well as learn implicit features that are able to further discriminate between confusing cases. Experimental evaluation is conducted on the original Mammographic Image Analysis Society (MIAS) and mini-MIAS databases, as well as our own Seoul National University Bundang Hospital digital mammographic database. It is shown that the proposed method outperforms comparable methods in terms of receiver operating characteristic (ROC) and precision-recall curves for detection of individual μCs and free-response receiver operating characteristic (FROC) curve for detection of clustered μCs. 相似文献