Sequences cloning clone cells. Medical search. Frequent questions

To clone a human CTR (hCTR), a BIN-67 cDNA library was screened using a cDNA probe from a porcine renal CTR (pCTR) that we recently cloned. (jci.org)
Transfection of this cDNA into COS cells resulted in expression of receptors with high affinity for salmon CT (Kd = approximately 0.44 nM) and for human CT (Kd = approximately 5.4 nM). (jci.org)
The cloned cDNA encodes a putative peptide of 490 amino acids with seven potential transmembrane domains. (jci.org)
We and other groups discovered the human IL-17F gene from a human EST sequence, a genomic DNA clone, and T-cell cDNA sequences in 2001 [ 3 , 8 , 9 ]. (hindawi.com)

For each isolated cell, they separately clone the two genes encoding the heavy and light chains that compose an antibody, and that vary in sequence from cell to cell. (the-scientist.com)
Once researchers have cloned the pairs of heavy- and light-chain genes naturally found in the cells, they can produce recombinant versions of each antibody in the lab. (the-scientist.com)
The issue with it is that there are a lot of steps for each cell to clone those antibody genes. (the-scientist.com)
Using PCR, researchers then amplify the heavy and light chain genes from antibodies of interest, clone them into expression vectors, and sequence them. (the-scientist.com)
But rather than cloning the antibody heavy- and light-chain genes from each cell, as in a typical expression cloning set-up, Connors's group distributed the isolated cells in 384-well plates, where they cultured them in the presence of feeder cells and signaling molecules that help to keep the B cells alive and also stimulate them to secrete antibodies. (the-scientist.com)
These include environmental DNA samples that have proven difficult to clone and synthetic genes encoding toxic products. (jcvi.org)
The precise pathophysiology of inherited single cell and multilineage cytopenias has not been elucidated despite the identification of many of the genes mutated in these disorders. (medscape.com)
In contrast, recombinant antibodies are monoclonal antibodies that are generated by cloning antibody genes into expression vectors and do not involve the use of hybridomas. (sigmaaldrich.com)
The gene is localized on the same chromosome at the distance of about 50 kb from telomeric sequences of IL-17A gene, and both genes are in a tail-to-tail orientation [ 3 ]. (hindawi.com)
These immune assays will be advanced by the development of standardized viral stocks, such as HIV-1 infectious molecular clones (IMC), that i) express a reporter gene, ii) are representative of globally diverse subtypes and iii) are engineered to easily exchange envelope (env) genes for expression of sequences of interest. (plos.org)
During B cell development, the antibody-coding genes are shuffled and pasted together by cellular enzymes in a coordinated, but imprecise, process known as VDJ recombination. (databasefootball.com)
Simply combining the various antibody genes together generates millions of different sequences, but the enzymes randomly insert or delete nucleotides at each of the gene junctions, leading to tens of billions of so-called "variable region" sequences, guaranteeing that no matter what pathogen an individual may encounter, he or she has likely made a B cell that can kill it. (databasefootball.com)

When genome editing induces a phenotypic alteration that is detectable by fluorescence (i.e., cell-surface expression of a target that can be non-lethally assessed with fluorescently labeled antibody), FACS provides a method of enriching edited cells 9 , narrowing the number of clones to propagate and analyze. (nature.com)
In this approach, researchers first collect antibody-producing cells from mice exposed to an antigen of interest and fuse them to myeloma cells, forming so-called hybridomas that can survive for a long time in culture. (the-scientist.com)
By screening single hybrid cells for those that produce the antibody of interest, researchers can develop a cell line generating virtually unlimited amounts of highly specific antibodies. (the-scientist.com)
This technique, commonly referred to as expression cloning, "is powerful because you're not limited-any B cell that you can isolate, in any context, you can make an antibody from," Wilson says. (the-scientist.com)
It would be extremely laborious to use this strategy to try to identify an antibody against an antigen of interest using the blood of a person without an active immune response, he says, because millions of memory B cells are present in the blood, each of which is uniquely specific for a different antigen. (the-scientist.com)
CHARTING THE COURSE: B cells grow in culture in individual wells over a layer of feeder cells, and culture media from cells is screened for antibody-binding specificity or neutralizing activity. (the-scientist.com)
The immortalization of antibody-producing cells from patients infected with HIV can be inefficient, and even when cells are successfully immortalized, some clones tend to be unstable. (the-scientist.com)
Polyclonal antibodies are produced as a result of the activation of numerous different antibody producing B cells in an animal. (sigmaaldrich.com)
With monoclonal antibody production, one B cell produces only one specific sequence of antibody to a given antigen. (sigmaaldrich.com)
Hence, a monoclonal antibody is considered a collection of identical antibodies secreted by a single B cell clone with specificity for only one antigenic epitope. (sigmaaldrich.com)
In traditional methods, monoclonal antibodies are produced by fusion of B cells with an immortal cell to produce a hybridoma that produces many copies of the exact same antibody. (sigmaaldrich.com)
With this technology, the problems of cell line drift, antibody expression variations, and antibody sequence mutations associated with classical hybridoma production and storage can be avoided. (sigmaaldrich.com)
Recombinant antibody production from most species consist of a heavy and a light chain and it is important to note that to accomplish maximum IgG production in mammalian cells, one must achieve a balanced expression of both heavy and light chains. (sigmaaldrich.com)
However, expression of these two chains on separate vectors is not an efficient way to generate monoclonal antibody producing cell lines. (sigmaaldrich.com)
For recombinant antibody production, stable cell lines such as the Chinese hamster ovary (CHO) cells, human embryonic kidney 293 (HEK293), or NS0 murine myeloma cells are more commonly employed. (sigmaaldrich.com)
However, in pemphigus vulgaris, the antibody variable region is sufficient to disrupt skin cell adhesion and cause blisters, so trying to suppress these unruly variable regions with a law-abiding constant region can't override this pathologic effect. (databasefootball.com)
The focus of our laboratory's research program over the last decade has been to determine the pathways by which autoantibodies arise and to identify what features of the antibody gene or protein sequence separate disease-causing autoantibodies from the normal antibodies that protect from infection. (databasefootball.com)

Generation and characterization of infectious molecular clones of transmitted/founder HIV-1 subtype C viruses. (harvard.edu)

The expression vectors are transfected into cells, and the resulting antibodies are purified and tested for activity. (the-scientist.com)

Expanded box below shows location of sequencing primers, restriction sites for subcloning, and placement of insertion site within the toxic minigene. (neb.com)

In the upstream step, delivery of genome editing machinery to the cell type of interest generates efficient and specific edits. (nature.com)
Therefore, the development of a method that allows screening of edited cells and minimizes cell manipulation and hands-on culturing would constitute a significant addition to the current genome engineering toolbox. (nature.com)
The method may also speed genome sequencing by eliminating the need for biological cloning. (jcvi.org)
In his 2012 book Regenesis , Harvard geneticist George Church proposes a different approach for cloning extinct animals whose genome has been sequenced. (nationalgeographic.com)
With that publication, "genome engineering of mammalian cells just took a big step forward," he says. (nationalgeographic.com)

Cell engineering through gene editing is fundamentally a two-step bioprocess. (nature.com)
corresponding to the copy number of each gene in each clone. (github.com)
CHO cells are most commonly used as a stable expression system and HEK293 cells are highly useful for transient gene expression where protein is harvested within a few days following DNA delivery. (sigmaaldrich.com)
The molecular sequence of this clone aligns with the gene accession number as a point of reference only. (origene.com)
However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. (origene.com)
Read-through transcription from the neighboring upstream gene (NME1) generates naturally-occurring transcripts (NME1-NME2) that encode a fusion protein comprised of sequence sharing identity with each individual gene product. (origene.com)

It also allows cell-free cloning of genomic DNA isolated from bacteria. (jcvi.org)

The development of immunoglobulin receptor repertoire sequencing methods makes it possible to perform repertoire-wide disease association studies of antigen receptor sequences. (elifesciences.org)
chain repertoires, by establishing a near-complete list of unique receptor chain sequences, or 'clonotypes', present in a sample. (elifesciences.org)
Cloning, characterization, and expression of a human calcitonin receptor from an ovarian carcinoma cell line. (jci.org)
Interestingly, the hCTR sequence is remotely related to the cAMP receptor of Dictyostelium discoideum (21% identical), but is not significantly related to other G protein-coupled receptor sequences now in the data bases. (jci.org)

Despite improvements in the CRISPR molecular toolbox, identifying and purifying properly edited clones remains slow, laborious, and low-yield. (nature.com)

Since the 1996 birth of Dolly the sheep, the world's first cloned mammal, scientists have greatly expanded and improved on cloning techniques. (nationalgeographic.com)

The kit also allows direct cloning from amplification reactions without purification, and works well whether or not the primers used in the PCR possess 5´-phosphate groups. (neb.com)
These antibodies can be cloned from any species using suitable oligonucleotide primers. (sigmaaldrich.com)

These results suggest that effects of β1 on sodium channel gating are dependent on the genetic background and signal transduction pathways present in the cell type used in heterologous expression and emphasize the importance of analyzing the effects of β1 in vivo . (jneurosci.org)
Using new tricks of genetic engineering, researchers could make adjustments to the DNA in the human cell so it matches the code of the Neanderthal. (nationalgeographic.com)
Genetic sequencing was performed on HIV strains from 29 of the 92 seropositive patients from the practices of three HCWs. (cdc.gov)

The amino acid sequence of the hCTR is 73% identical to the pCTR, although the hCTR contains an insert of 16 amino acids between transmembrane domain I and II. (jci.org)
Among the IL-17 cytokine family members, IL-17F shows the highest amino acid sequence homology (50%) to IL-17A, while only 10-30% sequence identity is seen between IL-17A and the other family members [ 10 ]. (hindawi.com)

Structurally, the IL-17 cytokine family members have no sequence similarity to any other known cytokine or other mammalian proteins [ 2 ]. (hindawi.com)
The four conserved cysteines in the C-terminal half of the IL-17F sequence are shown to form a cystine knot structural motif in the crystal structure, and, interestingly, this cystine knot structure is similar to a common structural motif seen in several growth factors, such as bone morphogenic proteins (BMPs), TGF- β , nerve growth factor (NGF), and platelet-derived growth factor (PDGF) [ 9 ]. (hindawi.com)
Our laboratory researches a rare and debilitating autoantibody-mediated disease called pemphigus vulgaris, in which the immune system generates antibodies against desmogleins, which are proteins responsible for holding skin cells together. (databasefootball.com)

β1 subunits affect sodium channel gating and cell surface expression when expressed in heterologous cells in vitro . (jneurosci.org)
Thus far, a subtype B IMC backbone expressing Renilla luciferase (LucR), and into which the ectodomain of heterologous env coding sequences can be expressed has been successfully developed but as execution of HIV-1 vaccine efficacy trials shifts increasingly to non-subtype B epidemics (Southern African and Southeast Asia), non-subtype B HIV-1 reagents are needed to support vaccine development. (plos.org)

And you can't mix your cells, because you'll get a heavy chain from one cell and a light chain from another cell," and therefore may not end up with a natural combination of heavy and light chains. (the-scientist.com)

This platform acquired its unique name because of its proprietary B cell immortalization technology that also allows us to produce monoclonal antibodies in a wide range of species. (sigmaaldrich.com)

Mechanochemical Principles of Spatial and Temporal Patterns in Cells and Tissues. (mpi-cbg.de)
Patterns are ubiquitous in living systems and underlie the dynamic organization of cells, tissues, and embryos. (mpi-cbg.de)

More recently, Wilson and others have devised ways to isolate single, antigen-specific B cells from the blood of human donors with active immune responses against a particular pathogen or vaccine. (the-scientist.com)
This technique is best for "when you have an enriched population of antigen-specific cells-they're not there, except for a short time after vaccination or infection," says Wilson. (the-scientist.com)
While previous researchers have obtained HIV-neutralizing antibodies using fluorescent probes to pull out antigen-specific memory B cells from a sample of blood, Connors and his team wanted to discover new HIV-neutralizing antibodies, without being limited only to those that are specific for known antigens. (the-scientist.com)

Sodium channel β1 subunits modulate α subunit gating and cell surface expression and participate in cell adhesive interactions in vitro . (jneurosci.org)

The downstream step involves identification and selection of the cells that have been properly edited. (nature.com)

Selection of edited cell clones currently relies on limiting dilution or fluorescence-activated cell sorting (FACS)-based single-cell sorting to isolate single cells. (nature.com)

Clone cells also include populations of recombinant DNA molecules all carrying the same inserted sequence. (harvard.edu)

As a consequence, investigators often need to isolate a large number of clones and then proceed with tedious and time-consuming efforts to expand all of them individually. (nature.com)

Vector species, T. cruzi infection status, collection location, number of clones sequenced, number and identity of taxa, and number of haplotypes represented in the clone sequences are indicated. (cdc.gov)
In 2003, researchers in Spain were the first to bring back an extinct species -the Pyrenean ibex, a wild mountain goat also called a bucardo-though the clone only lived for a few minutes. (nationalgeographic.com)
Starting with an intact cell (fresh or frozen) of the animal they'd like to clone, scientists first remove the nucleus, where DNA resides, and insert it into a hollowed-out egg cell of the same or a related species. (nationalgeographic.com)
It starts with a healthy cell of a closely related species-cloning a Neanderthal, for example, could start with a stem cell from a modern human. (nationalgeographic.com)

This procedure allows cell-free cloning of individual synthetic DNA molecules that cannot be cloned in Escherichia coli, for example synthetic phage genomes carrying lethal mutations. (jcvi.org)
Mutations in the CT have been shown to affect the conformation of gp120 ectodomain [ 16 - 18 ], and more recently, Durham et al suggested that the CT regulates the conformation of Env at the cell surface and control epitope exposure through T cell virological synapses [ 19 ]. (plos.org)

Clone Cells" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings) . (harvard.edu)

However, the effects of β1 are highly dependent on the experimental system in which they are studied, and different effects on the kinetics and voltage dependence of gating of brain sodium channels are observed on expression in Xenopus oocytes, Chinese hamster lung and ovary cells, and human embryonic kidney cells (Isom et al. (jneurosci.org)
assigns single-cell RNA-seq expression to cancer clones by probabilistically mapping RNA-seq to clone-specific copy number profiles using reparametrization gradient variational inference . (github.com)
Cell clusters were identified based on protein expression only. (biolegend.com)
The TotalSeq™-C Human TBNK Cocktail has been designed to react with immune cells as defined and classified by the expression of the surface antigens CD19, CD3, CD16, CD4, CD11c, CD56 (NCAM), CD14, CD8, CD45. (biolegend.com)
This clone was engineered to express the complete ORF with an expression tag. (origene.com)

A human ovarian small cell carcinoma line (BIN-67) expresses abundant calcitonin (CT) receptors (CTR) (143,000 per cell) that are coupled, to adenylate cyclase. (jci.org)
The dissociation constants (Kd) for the CTRs on these BIN-67 cells is approximately 0.42 nM for salmon CT and approximately 4.6 nM for human CT. (jci.org)
Human peripheral blood mononuclear cells were stained with the TotalSeq™-C TBNK Cocktail and processed using the 10x Genomics Single Cell 5' v1.1 feature barcoding kit and Illumina sequencing. (biolegend.com)
While cocktail is being centrifuged, block cells by adding 5 µL of Human TruStain FcX™ Fc blocking reagent (Cat. (biolegend.com)
The human immune system is a complex network of cells and soluble factors that can cleverly adapt to combat infection. (databasefootball.com)
If a human cell could be Neanderthalized, it would be implanted into the womb of a surrogate mother, either a woman or a chimp, and then develop into a fetus. (nationalgeographic.com)

Each colony is split and sequentially exported for on-target sequencing and further off-chip clonal expansion of the validated clones. (nature.com)
This PCR Cloning Kit contains an optimized Cloning Mix containing a proprietary ligation enhancer and a linearized vector that uses a novel mechanism for background colony suppression to give a low background. (neb.com)

These three cytokines are differentially expressed in activated CD4 + T cells. (hindawi.com)

We have recently identified, for the first time, that bronchial epithelial cells are its novel cell source in response to IL-33 via ST2-ERK1/2-MSK1 signaling pathway. (hindawi.com)

Moreover, even when high-purity single-cell sorting can be achieved, viability after sorting is often low to moderate, especially for cell types that are particularly sensitive to hydrodynamic stress or low-density culture conditions (e.g., primary cells or pluripotent stem cell lines). (nature.com)
The proliferation and differentiation of pluripotent stem cells give rise to progeny that can populate the entire immunologic and hematopoietic systems through committed progenitors of both the lymphoid and myeloid lineages. (medscape.com)

A group of genetically identical cells all descended from a single common ancestral cell by mitosis in eukaryotes or by binary fission in prokaryotes. (harvard.edu)

Ultraviolet radiation shapes dendritic cell leukaemia transformation in the skin. (harvard.edu)

Cell-free cloning using phi29 DNA polymerase. (jcvi.org)
Cell-free cloning of single circular molecules by using phi29 DNA polymerase was achieved by carrying out the amplification reactions in very small volumes, typically 600 nl. (jcvi.org)
This DNA can be sequenced directly from the phi29 DNA polymerase reaction without further amplification. (jcvi.org)
The ultimate in flexibility: clone with any amplicon made with any DNA polymerase, with or without 5´ phosphates, purified or not! (neb.com)

Also available without competent cells . (neb.com)

Accelerated hematopoietic cell apoptosis has been demonstrated in virtually all IBMFS. (medscape.com)
For example, B cells rearrange and mutate their own DNA to create a vast repertoire of antibodies capable of binding to and destroying virtually any target. (databasefootball.com)
Unfortunately, because B cells harbor a virtually unlimited library of sequences, they have the potential to create delinquent antibodies that react against self, rather than foreign invaders. (databasefootball.com)

After the B cell leaves the bone marrow, if it meets its target, it can activate, divide, and further diversify its variable region through somatic hypermutation of its DNA, spawning clans or lineages of related but distinct B cells all descending from a single precursor, optimally evolved to ward off their preferred invading pathogen. (databasefootball.com)

Clonal dynamics of alloreactive T cells in kidney allograft rejection after anti-PD-1 therapy. (harvard.edu)

Inherited bone marrow failure syndromes (IBMFS) are usually the result of intrinsic stem cell/progenitor defects. (medscape.com)
Colorimetric Barcoding to Track, Isolate, and Analyze Hematopoietic Stem Cell Clones. (harvard.edu)

These data illustrated that whereas CD47/SIRPα inhibition could potentiate tumor cell phagocytosis, CD40-mediated activation of a type I interferon response provided a bridge between macrophage- and T-cell-mediated immunity that significantly enhanced durable tumor control and rejection. (aacrjournals.org)

Progress has been made to enhance the efficiency of HDR-based editing 8 , however, a technology to identify cells with desired mono- or bi-allelic edits is urgently needed to realize the full potential of CRISPR. (nature.com)
Cell-free cloning has many potential applications to both natural and synthetic DNA. (jcvi.org)
Localized heterochrony integrates overgrowth potential of oncogenic clones. (harvard.edu)

Tumor immunogenicity results from the processing, and subsequent presentation of tumor antigens released from dead or dying tumor cells on MHC molecules expressed on antigen-presenting cells (APC). (aacrjournals.org)

The etiology of bone marrow failure (BMF) includes defective stem/progenitor cells and/or stroma/accessory cells/growth factors, as well as deficient nonspecific nutrients or, as in the case of acquired aplastic anemia, immune-mediated abnormalities. (medscape.com)

Hematopoietic cells are almost totally absent. (medscape.com)
The microenvironment, consisting of lymphocytes, macrophages, fibroblasts, endothelial elements, and stroma in which hematopoietic cells reside, creates a regulatory niche that determines the local area network. (medscape.com)

Here, we establish a method to enable clonal isolation, selection, and expansion of properly edited cells, using OptoElectroPositioning technology for single-cell manipulation on a nanofluidic device. (nature.com)
Briefly, after electroporation of primary T cells with CXCR4 -targeting Cas9 ribonucleoproteins, single T cells are isolated on a chip and expanded into colonies. (nature.com)
Using this method, single-clone editing efficiencies, including the rate of mono- and bi-allelic indels or precise nucleotide replacements, can be assessed within 10 days from Cas9 ribonucleoprotein introduction in cells. (nature.com)
This is particularly useful when clones have been inferred using ultra-shallow single-cell DNA-seq meaning SNV analysis is not possible. (github.com)
This reagent is a combination of TotalSeq™-C oligo conjugated clones at optimal concentrations for single-cell sequencing analysis. (biolegend.com)
These specializations can change through a process called class switching, in which a B cell shuffles the same variable region onto a different constant region, meaning that a single clan of B cells can express antibodies of many different isotypes and subclasses. (databasefootball.com)

Major immune cell lineages are identified using the TotalSeq™-C TBNK Cocktail. (biolegend.com)

We also offer solutions for automation, site-directed mutagenesis, as well as your favorite restriction enzyme, ligase or competent cell products. (neb.com)
In this study, we found that RING finger protein 4 (RNF4), a RING finger E3 ubiquitin ligase, is required for the RIPK1 autophosphorylation and subsequent cell death. (mdpi.com)

β1 also recruits ankyrin to the plasma membrane at sites of cell-cell contact in response to homophilic cell adhesion (Malhotra et al. (jneurosci.org)
The C121Wβ1 mutation also disrupted homophilic β1-β1 cell adhesion. (jneurosci.org)

It has been reported that RNF4 negatively regulates TNF-α-induced activation of the nuclear factor-κB (NF-κB) through downregulation of transforming growth factor β-activated kinase 1 (TAK1) activity, indicating the possibility that RNF4-mediated TAK1 suppression results in enhanced sensitivity to cell death. (mdpi.com)

IgA antibodies act as sentries at mucosal surfaces, IgE recruit in cells specialized for allergic reactions, and IgG are general-purpose antibodies with multiple functions, divided across 4 different subclasses (IgG1-IgG4). (databasefootball.com)
IgG1, IgG2, and IgG3 bind to receptors on inflammatory cells and recruit in mediators that can help to destroy targets, whereas IgG4 is relatively inert, which can be helpful for calming down inflammation by blocking other antibodies from binding and causing damage. (databasefootball.com)
Self-reactive antibodies (autoantibodies) are usually weeded out at "tolerance checkpoints" as the B cell matures, but these self-defense tactics are far from perfect. (databasefootball.com)

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