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tac Vector for High-Level Bacterial Expression. The Tac-Promoter (abbreviated as Ptac), or tac vector is a synthetically produced DNA promoter, produced from the combination of promoters from the trp and lac operons. [1] It is commonly used for protein production in Escherichia coli. [2] Two hybrid promoters functional in Escherichia coli were ...
To perform a run-off transcription assay, a gene of interest, including the promoter, is cloned into a plasmid. [4] The plasmid is digested at a known restriction enzyme cut site downstream from the transcription start site such that the expected mRNA run-off product would be easily separated by gel electrophoresis. [1] [2] [4]
The promoters used for these vector are usually based on the promoter of the lac operon or the T7 promoter, [11] and they are normally regulated by the lac operator. These promoters may also be hybrids of different promoters, for example, the Tac-Promoter is a hybrid of trp and lac promoters. [12]
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.
The lacUV5 promoter is a mutated promoter from the Escherichia coli lac operon which is used in molecular biology to drive gene expression on a plasmid. lacUV5 is very similar to the classical lac promoter, containing just 2 base pair mutations in the -10 hexamer region, compared to the lac promoter. [1]
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).
Vector map of pUC19. pUC19 is one of a series of plasmid cloning vectors designed by Joachim Messing and co-workers. [1] The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the abbreviation for the University of California, where early work on the plasmid series had been conducted. [2]
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 ( mRNA ), or can have a function in and of itself, such as tRNA or rRNA .