|
|
(14 intermediate revisions by 5 users not shown) |
Line 1: |
Line 1: |
− | [[Image:Part icon tag.png]]
| + | #redirect [[Help:Proteins]] |
− | <small>Browse [https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=Tag Tag parts]!</small>
| + | |
− | <hr>
| + | |
− | | + | |
− | [https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=Tag (Degradation) Tags] are genetic additions to the end of a sequence which mark a protein for degradation, thus decreasing a protein's half life.
| + | |
− | One of the useful aspects of genetic tags is the ability to detect gene activity in a time-sensitive manner.
| + | |
− | | + | |
− | Tags can be found on:
| + | |
− | * [[Image:Part icon reporter.png]][https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=reporter Reporters],
| + | |
− | * [[Image:Part icon cds.png]][https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=coding Protein Coding parts],
| + | |
− | * [[Image:Part icon cds.png]][https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=generator Protein Generators], and
| + | |
− | * [[Image:Part icon cds.png]][https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=inverter Inverters].<br>
| + | |
− | ''Note'': Tags cannot currently be used as [[An Introduction to BioBricks|Biobricks]] because currently all of our [https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=Coding Protein Coding parts] end in stop codons.
| + | |
− | | + | |
− | ==Mechanism of Use==
| + | |
− | Currently all of our [https://parts.igem.org/cgi/partsdb/pgroup.cgi?pgroup=Tag Tags] operate through the use of protein-degrading enzymes (proteases) within the cell.
| + | |
− | They do so by coding for a sequence of about eleven amino acids at the C-terminus of a protein. This sequence is read by a special type of RNA known as ssRA ("small stable RNA A"), and then degraded by the proteases ClpXP or ClpAP in e.coli (no such system is yet known for yeast).
| + | |
− | | + | |
− | Although originally the number of amino acids encoding for a ssRA tag was ''eleven'' (the first sequence that encoded for destruction via ssRA tagging was <b>AANDENYALAA</b> by [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8584937 Keiler et al]), a subsequent study by [http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=9603842 Andersen et al] tested the efficacy of mutating only the last ''three'' amino acids of that system. Thus our tags (<b>AAV, ASV, LVA, LAA</b>) are classified by only three amino acids.
| + | |
− | | + | |
− | | + | |
− | ==Future Plans==
| + | |
− | Our future plans for tags involve building proteins using biobricks. To read more on this project, visit [https://dspace.mit.edu/handle/1721.1/32535 Ira Phillip's work on biobrick modification].
| + | |
− | | + | |
− | | + | |
− | ==References==
| + | |
− | # Jabri and Nature News. "Tag, you're degraded". 2003. [http://www.nature.com/nsmb/journal/v10/n9/full/nsb0903-676.html article] | + | |
− | # Karzai, AW. "The SsrA−SmpB system for protein tagging, directed degradation and ribosome rescue". 2000. [http://www.nature.com/nsmb/journal/v7/n6/full/nsb0600_449.html Review Article]
| + | |
− | # Keiler, KC et al. "Role of a peptide tagging system in degradation of proteins synthesized from damaged messenger RNA." 1996. [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=8584937 Pubmed]
| + | |
− | # Andersen, JB et al. "New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria". 1998. [http://www.pubmedcentral.gov/articlerender.fcgi?tool=pubmed&pubmedid=9603842 link]
| + | |