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Bianca V Gapp Tomasz Konopka Thomas Penz Vineet Dalal Tilmann Bürckstümmer Christoph Bock Sebastian MB Nijman 《Molecular systems biology》2016,12(8)
Reverse genetic screens have driven gene annotation and target discovery in model organisms. However, many disease‐relevant genotypes and phenotypes cannot be studied in lower organisms. It is therefore essential to overcome technical hurdles associated with large‐scale reverse genetics in human cells. Here, we establish a reverse genetic approach based on highly robust and sensitive multiplexed RNA sequencing of mutant human cells. We conduct 10 parallel screens using a collection of engineered haploid isogenic cell lines with knockouts covering tyrosine kinases and identify known and unexpected effects on signaling pathways. Our study provides proof of concept for a scalable approach to link genotype to phenotype in human cells, which has broad applications. In particular, it clears the way for systematic phenotyping of still poorly characterized human genes and for systematic study of uncharacterized genomic features associated with human disease. 相似文献
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Evidence for arrested G2 cell subpopulation in the Ehrlich ascite carcinoma has been provided using flow cytometry stathmokinetic and bromdesoxyuridine--Hoechst 33258 techniques. After exposure in DME--F-12 media G2-arrested cells synchronically enter mitosis and then move trough G1 period. Durations have been measured for G1. G2 phases and G2-G1 transition. 相似文献
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Cell-to-cell fusion (cell fusion) is a fundamental biological process that also has been used as a versatile experimental tool to dissect a variety of cellular mechanisms, including the consequences of cell fusion itself, and to produce cells with desired properties, such as hybridomas and reprogrammed progenitors. However, current methods of cell fusion are not satisfactory because of their toxicity, inefficiency, or lack of flexibility. We describe a simple, versatile, scalable, and nontoxic approach that we call V-fusion, as it is based on the ability of the vesicular stomatitis virus G protein (VSV-G), a viral fusogen of broad tropism, to become rapidly and reversibly activated. We suggest that this approach will benefit a broad array of studies that investigate consequences of cell fusion or use cell fusion as an experimental tool. 相似文献
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Most estrous cycles in cows consist of 2 or 3 waves of follicular activity. Waves of ovarian follicular development comprise
the growth of dominant follicles some of which become ovulatory and the others are anovulatory. Ovarian follicular activity
in cows during estrous cycle was studied with a special reference to follicular waves and the circulating concentrations of
estradiol and progesterone. Transrectal ultrasound examination was carried out during 14 interovulatory intervals in 7 cows.
Ovarian follicular activity was recorded together with assessment of serum estradiol and progesterone concentrations. Three-wave
versus two-wave interovulatory intervals was observed in 71.4% of cows. The 3-wave interovulatory intervals differed from
2-wave intervals in: 1) earlier emergence of the dominant follicles, 2) longer in length, and 3) shorter interval from emergence
to ovulation. There was a progressive increase in follicular size and estradiol production during growth phase of each wave.
A drop in estradiol concentration was observed during the static phase of dominant anovulatory follicles. The size of the
ovulatory follicle was always greater and produced higher estradiol compared with the anovulatory follicle. In conclusion,
there was a predominance of 3-wave follicular activity that was associated with an increase in length of interovulatory intervals.
A dominant anovulatory follicle during its static phase may initiate the emergence of a subsequent wave. Follicular size and
estradiol concentration may have an important role in controlling follicular development and in determining whether an estrous
cycle will have 2 or 3-waves. 相似文献
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The O-antigen chain from the lipopolysaccharide of Helicobacter pylori
strain UA861 was determined to be composed of an elongated type 2 N -
acetyllactosamine backbone,
-[-->3)-beta-D-Gal-(1-->4)-beta-D-GlcNAc-(1- ]n-->, with
approximately half of the GlcNAc units carrying a terminal alpha-d-Glc
residue at the O -6 position. The O-chain of H.pylori UA861 was terminated
by a N -acetyllactosamine [beta-D-Gal-(1-->4)-beta-D- GlcNAc] (LacNAc)
epitope and did not express terminal Lewis X or Lewis Y blood-group
determinants as previously found in other H.pylori strains. The absence of
terminal Lewis X and Lewis Y blood-group epitopes and the replacement of
Fuc by Glc as a side chain in the O- chain of H.pylori UA861 represents yet
another type of lipopolysaccharide structure from H.pylori species. These
structural differences in H.pylori lipopolysaccharide molecules carry
implications with regard to possible different pathogenic events between
strains and respective hosts.
相似文献