MRNA Display
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MRNA Display
mRNA display is a display technique used for ''in vitro'' protein, and/or peptide evolution to create molecules that can bind to a desired target. The process results in translated peptides or proteins that are associated with their mRNA progenitor via a puromycin linkage. The complex then binds to an immobilized target in a selection step (affinity chromatography). The mRNA-protein fusions that bind well are then reverse transcribed to cDNA and their sequence amplified via a polymerase chain reaction. The result is a nucleotide sequence that encodes a peptide with high affinity for the molecule of interest. Puromycin is an analogue of the 3’ end of a tyrosyl-tRNA with a part of its structure mimics a molecule of adenosine, and the other part mimics a molecule of tyrosine. Compared to the cleavable ester bond in a tyrosyl-tRNA, puromycin has a non-hydrolysable amide bond. As a result, puromycin interferes with translation, and causes premature release of translation products. ...
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In Vitro
''In vitro'' (meaning in glass, or ''in the glass'') studies are performed with microorganisms, cells, or biological molecules outside their normal biological context. Colloquially called " test-tube experiments", these studies in biology and its subdisciplines are traditionally done in labware such as test tubes, flasks, Petri dishes, and microtiter plates. Studies conducted using components of an organism that have been isolated from their usual biological surroundings permit a more detailed or more convenient analysis than can be done with whole organisms; however, results obtained from ''in vitro'' experiments may not fully or accurately predict the effects on a whole organism. In contrast to ''in vitro'' experiments, '' in vivo'' studies are those conducted in living organisms, including humans, and whole plants. Definition ''In vitro'' ( la, in glass; often not italicized in English usage) studies are conducted using components of an organism that have been isolated ...
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Nucleic Acids Research
''Nucleic Acids Research'' is an open-access peer-reviewed scientific journal published since 1974 by the Oxford University Press. The journal covers research on nucleic acids, such as DNA and RNA, and related work. According to the ''Journal Citation Reports'', the journal's 2021 impact factor is 19.160. The journal publishes two yearly special issues, the first issue of each year is dedicated to biological databases, published in January since 1993, and the other is devoted to papers describing web-based software resources of value to the biological community (web server A web server is computer software and underlying hardware that accepts requests via HTTP (the network protocol created to distribute web content) or its secure variant HTTPS. A user agent, commonly a web browser or web crawler, initia ...s), published in July since 2003. References External links * * Archives , https://www.ncbi.nlm.nih.gov/pmc/journals/4/ Biochemistry journals Open access ...
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Current Opinion In Chemical Biology
Currents, Current or The Current may refer to: Science and technology * Current (fluid), the flow of a liquid or a gas ** Air current, a flow of air ** Ocean current, a current in the ocean *** Rip current, a kind of water current ** Current (stream), currents in rivers and streams ** Convection current, flow caused by unstable density variation due to temperature differences * Current (mathematics), geometrical current in differential topology * Conserved current, a field associated to a symmetry in field theory * Electric current, a flow of electric charge through a medium * Probability current, in quantum mechanics * IBM Current, an early personal information management program Arts and entertainment Music * ''Current'' (album), a 1982 album by Heatwave * ''Currents'' (Eisley album) * ''Currents'' (Tame Impala album) * "The Current" (song), by the Blue Man Group * "Currents", a song by Dashboard Confessional from ''Dusk and Summer'', 2006 * "Currents", a song by Drake fr ...
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Ribosome Display
Ribosome display is a technique used to perform ''in vitro'' protein evolution to create proteins that can bind to a desired ligand. The process results in translated proteins that are associated with their mRNA progenitor which is used, as a complex, to bind to an immobilized ligand in a selection step. The mRNA-protein hybrids that bind well are then reverse transcribed to cDNA and their sequence amplified via PCR. The result is a nucleotide sequence that can be used to create tightly binding proteins. Ribosome display process Ribosome display begins with a native library of DNA sequences coding for polypeptides. Each sequence is transcribed, and then translated ''in vitro'' into polypeptide. However, the DNA library coding for a particular library of binding proteins is genetically fused to a spacer sequence lacking a stop codon before its end. The lack of a stop codon prevents release factors from binding and triggering the disassembly of the translational complex. So, thi ...
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Yeast Display
Yeast display (or yeast surface display) is a protein engineering technique that uses the expression of recombinant proteins incorporated into the cell wall of yeast for isolating and engineering antibodies. Development The yeast display technique was first published by the laboratory of Professor K. Dane Wittrup and Eric T. Boder. The technology was sold to Abbott Laboratories in 2001. How it works A protein of interest is displayed as a fusion to the Aga2p protein on the surface of yeast. The Aga2p protein is naturally used by yeast to mediate cell–cell contacts during yeast cell mating. As such, display of a protein via Aga2p projects the protein away from the cell surface, minimizing potential interactions with other molecules on the yeast cell wall. The use of magnetic separation and flow cytometry in conjunction with a yeast display library is a highly effective method to isolate high affinity protein ligands against nearly any receptor through directed evolution. Advant ...
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Bacterial Display
Bacterial display (or bacteria display or bacterial surface display) is a protein engineering technique used for in vitro protein evolution. Libraries of polypeptides displayed on the surface of bacteria can be screened using flow cytometry or iterative selection procedures (biopanning). This protein engineering technique allows us to link the function of a protein with the gene that encodes it. Bacterial display can be used to find target proteins with desired properties and can be used to make affinity ligands which are cell-specific. This system can be used in many applications including the creation of novel vaccines, the identification of enzyme substrates and finding the affinity of a ligand for its target protein. Bacterial display is often coupled with magnetic-activated cell sorting (MACS) or fluorescence-activated cell sorting (FACS) techniques. Competing methods for protein evolution ''in vitro'' are phage display, ribosome display, yeast display, and mRNA display. Ba ...
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Phage Display
Phage display is a laboratory technique for the study of protein–protein, protein–peptide, and protein– DNA interactions that uses bacteriophages (viruses that infect bacteria) to connect proteins with the genetic information that encodes them. In this technique, a gene encoding a protein of interest is inserted into a phage coat protein gene, causing the phage to "display" the protein on its outside while containing the gene for the protein on its inside, resulting in a connection between genotype and phenotype. These displaying phages can then be screened against other proteins, peptides or DNA sequences, in order to detect interaction between the displayed protein and those other molecules. In this way, large libraries of proteins can be screened and amplified in a process called ''in vitro'' selection, which is analogous to natural selection. The most common bacteriophages used in phage display are M13 and fd filamentous phage, though T4, T7, and λ phage have ...
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ACS Combinatorial Science
''ACS Combinatorial Science'' (usually abbreviated as ''ACS Comb. Sci.''), formerly ''Journal of Combinatorial Chemistry'' (1999-2010), was a peer-reviewed scientific journal, published since 1999 by the American Chemical Society. ''ACS Combinatorial Science'' publishes articles, reviews, perspectives, accounts and reports in the field of Combinatorial Chemistry. ''JCS'' is currently indexed in: Chemical Abstracts Service (CAS), SCOPUS, EBSCOhost, PubMed, and Web of Science. Anthony Czarnik } Anthony W. Czarnik (born 1957) is an American chemist and inventor. He is best known for pioneering studies in the field of fluorescent chemosensors and co-founding Illumina, Inc., a biotechnology company in San Diego. Czarnik was also the fou ... served the founding editor; he served as Editor from 1999-2010. M.G. Finn served as Editor from 2010-2020. In 2010, ACS agreed to change the name of the journal to ''"Combinatorial Science"'' and it was the first and only ACS journal to be dev ...
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Reverse Transcriptase
A reverse transcriptase (RT) is an enzyme used to generate complementary DNA (cDNA) from an RNA template, a process termed reverse transcription. Reverse transcriptases are used by viruses such as HIV and hepatitis B to replicate their genomes, by retrotransposon mobile genetic elements to proliferate within the host genome, and by eukaryotic cells to extend the telomeres at the ends of their linear chromosomes. Contrary to a widely held belief, the process does not violate the flows of genetic information as described by the classical central dogma, as transfers of information from RNA to DNA are explicitly held possible. Retroviral RT has three sequential biochemical activities: RNA-dependent DNA polymerase activity, ribonuclease H (RNase H), and DNA-dependent DNA polymerase activity. Collectively, these activities enable the enzyme to convert single-stranded RNA into double-stranded cDNA. In retroviruses and retrotransposons, this cDNA can then integrate into the host ...
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Chemistry & Biology
''Cell Chemical Biology'' is a monthly, peer-reviewed, scientific journal published by Cell Press. The journal publishes research in chemical biology and studies at the interface of chemistry and biology Biology is the scientific study of life. It is a natural science with a broad scope but has several unifying themes that tie it together as a single, coherent field. For instance, all organisms are made up of cells that process hereditary .... References External links * Biochemistry journals Cell Press academic journals Monthly journals {{biochemistry-journal-stub ...
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Figure 3
Figure may refer to: General *A shape, drawing, depiction, or geometric configuration * Figure (wood), wood appearance *Figure (music), distinguished from musical motif *Noise figure, in telecommunication * Dance figure, an elementary dance pattern *A person's figure, human physical appearance Arts *Figurine, a miniature statuette representation of a creature *Action figure, a posable jointed solid plastic character figurine *Figure painting, realistic representation, especially of the human form *Figure drawing * Model figure, a scale model of a creature Writing *figure, in writing, a type of floating block (text, table, or graphic separate from the main text) *Figure of speech, also called a rhetorical figure *Christ figure, a type of character * in typesetting, text figures and lining figures Accounting *Figure, a synonym for number A number is a mathematical object used to count, measure, and label. The original examples are the natural numbers 1, 2, 3, 4, a ...
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DNA Ligase
DNA ligase is a specific type of enzyme, a ligase, () that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms, but some forms (such as DNA ligase IV) may specifically repair double-strand breaks (i.e. a break in both complementary strands of DNA). Single-strand breaks are repaired by DNA ligase using the complementary strand of the double helix as a template, with DNA ligase creating the final phosphodiester bond to fully repair the DNA. DNA ligase is used in both DNA repair and DNA replication (see '' Mammalian ligases''). In addition, DNA ligase has extensive use in molecular biology laboratories for recombinant DNA experiments (see '' Research applications''). Purified DNA ligase is used in gene cloning to join DNA molecules together to form recombinant DNA. Enzymatic mechanism The mechanism of DNA ligase is to form two covalent p ...
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