scholarly journals Evolution of Avian coronavirus (AvCoV) in BHK-21 and VERO cells

2020 ◽  
Vol 57 (2) ◽  
pp. e166086
Author(s):  
Paulo Eduardo Brandão ◽  
Beatriz Alcântara Leite ◽  
Sueli Akemi Taniwaki Miyagi
Keyword(s):  
Cell Lines ◽  
Nonstructural Protein ◽  
Protein S ◽  
Vero Cells ◽  
Gradual Transition ◽  
Chicken Embryos ◽  
The Stability ◽  
Quasispecies Evolution ◽  
Selection Of

Avian coronavirus (AvCoV) infects a range of tissues in chickens and several other avian species. Although the virus can be isolated in chicken embryos, only a few strains of the 6 genotypes/33 lineages can grow in cell lines, with the Beaudette strain (GI-1 lineage) being the most used for in vitro studies. Considering the differences between cell lines and chicken embryos as habitats for AvCoV, this study aimed to assess the diversity of the genes coding for the nonstructural protein 3 (nsp3) and spike envelope protein (S) after serial passages in BHK-21 and Vero cells. After 14 passages of an embryo-adapted Beaudette strain, the virus loads fluctuated in both cell lines, with the highest loads being 8.72 log genome copies/µL for Vero and 6.36 log genome copies/µL for BHK-21 cells. No polymorphisms were found for nsp3; regarding S, not only aa substitutions (Vero: 8th passage A150S, and 14th S150A; BHK-21: 4th S53F, 8th F53Y, and 8th S95R), but also minor variants could be detected on chromatograms with fluctuating intensities. As the regions of these aa substitutions are within the receptor-binding domain of S, it can be speculated that differences in cell receptors between Vero and BHK-21 cells and the speed of cell death led to the selection of different dominant strains, while the stability of nsp3 supports its function as a protease involved in AvCoV replication. In conclusion, AvCoV quasispecies evolution is influenced by the biological model under consideration, and a gradual transition is seen for minor and major variants.

Protein synthesis is required for rapid degradation of tubulin mRNA and other deflagellation-induced RNAs in Chlamydomonas reinhardi

1986 ◽  
Vol 6 (1) ◽  
pp. 54-61
Author(s):  
E J Baker ◽  
L R Keller ◽  
J A Schloss ◽  
J L Rosenbaum
Keyword(s):  
Protein Synthesis ◽  
Transcriptional Control ◽  
Protein S ◽  
Protein Synthesis Inhibitors ◽  
Control Factors ◽  
Flagellar Proteins ◽  
The Stability ◽  
And Control

After flagellar detachment in Chlamydomonas reinhardi, there is a rapid synthesis and accumulation of mRNAs for tubulin and other flagellar proteins. Maximum levels of these mRNAs (flagellar RNAs) are reached within 1 h after deflagellation, after which they are rapidly degraded to their predeflagellation levels. The degradation of alpha- and beta-tubulin RNAs was shown to be due to the shortening of their half-lives after accumulation (Baker et al., J. Cell Biol. 99:2074-2081, 1984). Deflagellation in the presence of protein synthesis inhibitors results in the accumulation of tubulin and other flagellar mRNAs by kinetics similar to those of controls. However, unlike controls, in which the accumulated mRNAs are rapidly degraded, these mRNAs are stabilized in cycloheximide. The stabilization by cycloheximide is specific for the flagellar mRNAs accumulated after deflagellation, since there is no change in the levels of flagellar mRNAs in nondeflagellated (uninduced) cells in the presence of cycloheximide. The kinetics of flagellar mRNA synthesis after deflagellation are shown to be the same in cycloheximide-treated and control cells by in vivo labeling and in vitro nuclear runoff experiments. These results show that protein synthesis is not required for the induced synthesis of flagellar mRNAs, and that all necessary transcriptional control factors are present in the cell before deflagellation, but that protein synthesis is required for the accelerated degradation of the accumulated flagellar mRNAs. Since cycloheximide prevents the induced synthesis and accumulation of flagellar proteins, it is possible that the product(s) of protein synthesis required for the accelerated decay of these mRNAs is a flagellar protein(s). The possibility that one or more flagellar proteins autoregulate the stability of the flagellar mRNAs is discussed.

scholarly journals Transgenic human ES and iPS reporter cell lines for identification and selection of pluripotent stem cells in vitro

2014 ◽  
Vol 13 (2) ◽  
pp. 251-261 ◽  
Author(s):  
Dmitry A. Ovchinnikov ◽  
Drew M. Titmarsh ◽  
Patrick R.J. Fortuna ◽  
Alejandro Hidalgo ◽  
Samah Alharbi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Lines ◽  
Pluripotent Stem Cells ◽  
Reporter Cell ◽  
Selection Of

MAGNETIC SELECTION OF TRANSDUCED CELLS USING NOVEL SURFACE MARKERS

2021 ◽  
Vol 1 (19) ◽  
pp. 194-196
Author(s):  
O.F Kandarakov ◽  
A.V. Bruter ◽  
A.V. Petrovskaya ◽  
A.V. Belyavsky
Keyword(s):  
Magnetic Separation ◽  
Cell Lines ◽  
Surface Protein ◽  
Cell Selection ◽  
Surface Markers ◽  
Efficiency Of Selection ◽  
Selection Of

The possibility of using HA- and FLAG–tags embedded into CD52 surface protein for magnetic separation of transduced cells in vitro was investigated. The efficiency of selection of transfected cell lines, both with single and binary tags, was shown to exceed 85%. Thus, surface markers on the basis of CD52 protein with integrated HA- and FLAG-tags are applicable for cell selection by the MACS method.

Mouse thymic epithelial cell lines expressing “Aire” and peripheral tissue-specific antigens reproduce in vitro negative selection of T cells

2011 ◽  
Vol 317 (14) ◽  
pp. 2019-2030 ◽  
Author(s):  
Yoshitaka Yamaguchi ◽  
Atsushi Takayanagi ◽  
Jiabing Chen ◽  
Kosuke Sakai ◽  
Jun Kudoh ◽  
...  
Keyword(s):  
T Cells ◽  
Epithelial Cell ◽  
Cell Lines ◽  
Negative Selection ◽  
Thymic Epithelial Cell ◽  
Tissue Specific ◽  
Epithelial Cell Lines ◽  
Specific Antigens ◽  
Selection Of

In vitro selection of salt-tolerant cell lines in kallar grass [Diplachne fusca (L.) Beauv.]

2003 ◽  
Vol 3 (1) ◽  
pp. 49-52 ◽  
Author(s):  
MALEE NANAKORN ◽  
WALAIKARN JIAMJETJAROON ◽  
SRISOM SUWANAWONG ◽  
CHALERMCHAI WONGWATTANA ◽  
IE SUNG SHIM
Keyword(s):  
Cell Lines ◽  
In Vitro Selection ◽  
Salt Tolerant ◽  
Kallar Grass ◽  
Tolerant Cell ◽  
Selection Of

scholarly journals The Selection of NFκB Inhibitors to Block Inflammation and Induce Sensitisation to FasL-Induced Apoptosis in HNSCC Cell Lines Is Critical for Their Use as a Prospective Cancer Therapy

2019 ◽  
Vol 20 (6) ◽  
pp. 1306 ◽  
Author(s):  
Mario Scheurer ◽  
Roman Brands ◽  
Mohamed El-Mesery ◽  
Stefan Hartmann ◽  
Urs Müller-Richter ◽  
...  
Keyword(s):  
Cancer Therapy ◽  
Cell Lines ◽  
In Vitro Study ◽  
Keratinocyte Cell Line Hacat ◽  
Keratinocyte Cell ◽  
Nfκb Pathway ◽  
Induced Apoptosis ◽  
Selection Of ◽  
Hnscc Cell

Inflammation is a central aspect of tumour biology and can contribute significantly to both the origination and progression of tumours. The NFκB pathway is one of the most important signal transduction pathways in inflammation and is, therefore, an excellent target for cancer therapy. In this work, we examined the influence of four NFκB inhibitors—Cortisol, MLN4924, QNZ and TPCA1—on proliferation, inflammation and sensitisation to apoptosis mediated by the death ligand FasL in the HNSCC cell lines PCI1, PCI9, PCI13, PCI52 and SCC25 and in the human dermal keratinocyte cell line HaCaT. We found that the selection of the inhibitor is critical to ensure that cells do not respond by inducing counteracting activities in the context of cancer therapy, e.g., the extreme IL-8 induction mediated by MLN4924 or FasL resistance mediated by Cortisol. However, TPCA1 was qualified by this in vitro study as an excellent therapeutic mediator in HNSCC by four positive qualities: (1) proliferation was inhibited at low μM-range concentrations; (2) TNFα-induced IL-8 secretion was blocked; (3) HNSCC cells were sensitized to TNFα-induced cell death; and (4) FasL-mediated apoptosis was not disrupted.

In vitro selection of salt tolerant cell lines in Solanum tuberosum L.

2007 ◽  
Vol 51 (4) ◽  
pp. 728-734 ◽  
Author(s):  
F. Queiros ◽  
F. Fidalgo ◽  
I. Santos ◽  
R. Salema
Keyword(s):  
Solanum Tuberosum ◽  
Cell Lines ◽  
In Vitro Selection ◽  
Solanum Tuberosum L ◽  
Salt Tolerant ◽  
Tolerant Cell ◽  
Selection Of

scholarly journals Attachment and invasion ofToxoplasma gondiiandNeospora caninumto epithelial and fibroblast cell linesin vitro

Parasitology ◽  
2005 ◽  
Vol 131 (5) ◽  
pp. 583-590 ◽  
Author(s):  
YING LEI ◽  
M. DAVEY ◽  
J. T. ELLIS
Keyword(s):  
Cell Lines ◽  
Neospora Caninum ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Vero Cells ◽  
Host Cells ◽  
Trypsin Treatment ◽  
Epithelial Cell Lines ◽  
Pre Treatment

Attachment and invasion ofToxoplasma gondiiandNeospora caninumto a cat and a dog fibroblast cell line and 2 epithelial cell lines (a cat kidney and Vero) were comparedin vitrousing fluorescence antibody methodology. In addition, trypsin treatment of tachyzoites was used to determine whether protein molecules were essential to the process of invasion. The results show that bothT. gondiiandN. caninuminvaded all 4 cell lines, and that pre-treatment ofT. gondiitachyzoites with trypsin caused an increase in the ability of the parasite to invade these host cells. FurthermoreT. gondii, in comparison toN. caninum, invaded all 4 cell lines at greater levels. The results here support the conclusion that bothT. gondiiandN. caninumhave the ability to invade a variety of cell types including both dog and cat cells, and questions the utility of Vero cells as an appropriate host cell forin vitrostudies on the biology of these taxa.

scholarly journals Opposing Effects of Polyadenylation on the Stability of Edited and Unedited Mitochondrial RNAs in Trypanosoma brucei

2005 ◽  
Vol 25 (5) ◽  
pp. 1634-1644 ◽  
Author(s):  
Chia-Ying Kao ◽  
Laurie K. Read
Keyword(s):  
Trypanosoma Brucei ◽  
Rna Stability ◽  
Protein S ◽  
Rna Degradation ◽  
Nucleotide Composition ◽  
Rna Turnover ◽  
Degradation Reactions ◽  
The Stability ◽  
Opposing Effects

ABSTRACT Mitochondrial RNAs in Trypanosoma brucei undergo posttranscriptional RNA editing and polyadenylation. We previously showed that polyadenylation stimulates turnover of unedited RNAs. Here, we investigated the role of polyadenylation in decay of edited RPS12 RNA. In in vitro turnover assays, nonadenylated fully edited RNA degrades significantly faster than its unedited counterpart. Rapid turnover of nonadenylated RNA is facilitated by editing at just six editing sites. Surprisingly, in direct contrast to unedited RNA, turnover of fully edited RNA is dramatically slowed by addition of a poly(A)20 tail. The same minimal edited sequence that stimulates decay of nonadenylated RNA is sufficient to switch the poly(A) tail from a destabilizing to a stabilizing element. Both nucleotide composition and length of the 3′ extension are important for stabilization of edited RNA. Titration of poly(A) into RNA degradation reactions has no effect on turnover of polyadenylated edited RNA. These results suggest the presence of a protective protein(s) that simultaneously recognizes the poly(A) tail and small edited element and which blocks the action of a 3′-5′ exonuclease. This study provides the first evidence for opposing effects of polyadenylation on RNA stability within a single organelle and suggests a novel and unique regulation of RNA turnover in this system.

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