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Template:Team:Bonn:NetworkData
From 2013.igem.org
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| - | content.titleShort = " | + | content.titleShort = "Gene knock-out and knock-in"; |
content.titleLong = ""; | content.titleLong = ""; | ||
| - | content.summary= " | + | content.summary= "Another irreversible method of protein regulation in bacteria is the Gene knock in or –out. Here the genomic DNA is manipulated to get a gain or a loss of function, in other words the production or "not-production" of a protein. |
| - | content.text= ""; | + | content.text= "To insert the gene of interest in the bacteria genome Recombineering is often used. Utilising this system, it is possible to generate inserts or deletion. It is based on the homologous recombination of a small engineered DNA parts with a target gene. Recombination then occurs in the defined region of the genome, resulting in the insertion of this foreign sequence.</br>One way of doing it is by using bacteriophage proteins, from bacteriophage lambda, which mediates insertion of DNA cassettes <sup> <a href=#5.1>[5.1]</a> </sup> <sup> <a href=#5.2>[5.2]</a> </sup>.</br> Gene expression, so protein production, after a knock in can be regulated by a promoter. </br> The disadvantage of this way of protein regulation is obviously the durance it needs to activate the expression of the gene till the full appearance of the protein. </br>A knock out causes a full loss of function. When a genes sequence is interrupted, it may still be translated, but the resulting protein will be non-functional.Here it is not possible to regulate the protein activity. The protein misses at any time, so a control of activity levels is not possible. </br></br> <p><a name=5.1>[5.1] </a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC107131/ > Kenan Murphy, Use of bacteriophage &lambda recombination functions to promote gene replacement in Escherichia coli (1998) </a> </p> <p> <a name=5.2> [5.2] </a> <a href=http://www.ncbi.nlm.nih.gov/pmc/articles/PMC148353/> Rapid modification of bacterial artificial chromosomes by ET- recombination (1998) </a> </p>"; ; |
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Revision as of 22:34, 1 October 2013
