About: Promoter bashing

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Promoter bashing is a technique used in molecular biology to identify how certain regions of a DNA strand, commonly promoters, affect the transcription of downstream genes. Under normal circumstances, proteins bind to the promoter and activate or repress transcription. In a promoter bashing assay, specific point mutations or deletions are made in specific regions of the promoter and the transcription of the gene is then measured. The contribution of a region of the promoter can be observed by the level of transcription. If a mutation or deletion changes the level of transcription, then it is known that that region of the promoter may be a binding site or other regulatory element.

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rdf:type	topical concept yago:WikicatMolecularBiologyTechniques yago:Ability105616246 yago:Abstraction100002137 yago:Cognition100023271 yago:Know-how105616786 yago:Method105660268 yago:PsychologicalFeature100023100 yago:Technique105665146
rdfs:label	Promoter bashing (en)
rdfs:comment	Promoter bashing is a technique used in molecular biology to identify how certain regions of a DNA strand, commonly promoters, affect the transcription of downstream genes. Under normal circumstances, proteins bind to the promoter and activate or repress transcription. In a promoter bashing assay, specific point mutations or deletions are made in specific regions of the promoter and the transcription of the gene is then measured. The contribution of a region of the promoter can be observed by the level of transcription. If a mutation or deletion changes the level of transcription, then it is known that that region of the promoter may be a binding site or other regulatory element. (en)
foaf:depiction
dcterms:subject	Molecular genetics Molecular biology techniques
Wikipage page ID	16185953 (xsd:integer)
Wikipage revision ID	1123483898 (xsd:integer)
Link from a Wikipage to another Wikipage	Electrophoretic mobility shift assay DNA footprinting Polymerase chain reaction Molecular genetics Gel electrophoresis Gene expression Deletion (genetics) Transcription (biology) DNA Sequencing Molecular biology techniques Directionality (molecular biology) Molecular cloning Restriction digest Protein Reporter gene Assay DNA strand Molecular biology Site-directed mutagenesis Point mutations Promoter (biology) Restriction digestion
sameAs	Promoter bashing Promoter bashing Promoter bashing Promoter bashing
dbp:wikiPageUsesTemplate	dbt:Reflist dbt:Molecular_genetics_methods
thumbnail	wiki-commons:Special:FilePath/Promoter_bashing.svg?width=300
has abstract	Promoter bashing is a technique used in molecular biology to identify how certain regions of a DNA strand, commonly promoters, affect the transcription of downstream genes. Under normal circumstances, proteins bind to the promoter and activate or repress transcription. In a promoter bashing assay, specific point mutations or deletions are made in specific regions of the promoter and the transcription of the gene is then measured. The contribution of a region of the promoter can be observed by the level of transcription. If a mutation or deletion changes the level of transcription, then it is known that that region of the promoter may be a binding site or other regulatory element. Promoter bashing is often done with deletions from either the 5' or 3' end of the DNA strand; this assay is easier to perform based on repeated restriction digestion and gel-purifying fragments of specific sizes. It is often easiest to ligate the promoter into the reporter, generate a large amount of the reporter construct using PCR or growth in bacteria, and then perform serial restriction digests on this sample. The ability of upstream promoters can be easily assayed by removing segments from the 5' end, and the same for the 3' end of the strand for downstream promoters. As the promoter commonly contains binding sequences for proteins affecting transcription, those proteins are also necessary when testing the effects of the promoter. Proteins which associate with the promoter can be identified using an electrophoretic mobility shift assay (EMSA), and the effects of inclusion or exclusion of the proteins with the mutagenized promoters can be assessed in the assay. This allows the use of promoter bashing to not only discover the location on the DNA strand which affects transcription, but also the proteins which affect that strand. The effects of protein interactions with each other as well as the binding sites can also be assayed in this way; candidate proteins must instead be identified by protein/protein interaction assays instead of an EMSA. (en)
gold:hypernym	Technique
prov:wasDerivedFrom	wikipedia-en:Promoter_bashing?oldid=1123483898&ns=0
page length (characters) of wiki page	8146 (xsd:nonNegativeInteger)
foaf:isPrimaryTopicOf	wikipedia-en:Promoter_bashing
is Link from a Wikipage to another Wikipage of	Genetic engineering Promoter (genetics)
is foaf:primaryTopic of	wikipedia-en:Promoter_bashing

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