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An inducer functions in two ways; namely: By disabling repressors. The gene is expressed because an inducer binds to the repressor. The binding of the inducer to the repressor prevents the repressor from binding to the operator. RNA polymerase can then begin to transcribe operon genes. By binding to activators.
In genetics, a promoter is a sequence of DNA to which proteins bind to initiate transcription of a single RNA transcript from the DNA downstream of the promoter. The RNA transcript may encode a protein , or can have a function in and of itself, such as tRNA or rRNA.
The tac promoter consists of the '–35' region of the trp promoter and the '–10' region of the lac promoter (and differs from a related trc promoter by 1 bp [3]). The tac promoter is, therefore, inducible by IPTG (Isopropyl β- D -1-thiogalactopyranoside), whilst also allowing higher maximum gene expression than either the lac or trp promoters.
Like allolactose, IPTG binds to the lac repressor and releases the tetrameric repressor from the lac operator in an allosteric manner, thereby allowing the transcription of genes in the lac operon, such as the gene coding for beta-galactosidase, a hydrolase enzyme that catalyzes the hydrolysis of β-galactosides into monosaccharides.
Inducible systems - An inducible system is off unless there is the presence of some molecule (called an inducer) that allows for gene expression. The molecule is said to "induce expression". The manner by which this happens is dependent on the control mechanisms as well as differences between prokaryotic and eukaryotic cells.
Example of a T-REx system controlling the expression of shRNA. Tetracycline-controlled transcriptional activation is a method of inducible gene expression where transcription is reversibly turned on or off in the presence of the antibiotic tetracycline or one of its derivatives (e.g. doxycycline).
Promoter - commonly used inducible promoters are promoters derived from lac operon and the T7 promoter. Other strong promoters used include Trp promoter and Tac-Promoter, which are a hybrid of both the Trp and Lac Operon promoters. Ribosome binding site (RBS) - follows the promoter, and promotes efficient translation of the protein of interest.
Methods to study promoter activity commonly are based in the expression of a reporter gene from the promoter of the gene of interest. [16] [2] [17] Mutations and deletions are made in a promoter region, and their changes on couple expression of the reporter gene are measured. [18] The most important reporter genes are the fluorescence proteins ...