What is pGEX 2t plasmid?
Bacterial vector for expressing GST fusion proteins with a thrombin site.
What is pGEX plasmid?
Plasmid: pGEX-1 Expression vector permitting production of a fusion protein. (ATCC staff) The glutathione S-transferase (GST) fusion protein can be purified by glutathione affinity chromatography, and the desired polypeptide released from the fusion product by thrombin.
What is a pGEX vector?
pGEX-2TK is designed to allow the detection of expressed proteins by directly labeling the tagged products in vitro. This vector contains the recognition sequence for the catalytic subunit of cAMP-dependent protein kinase obtained from heart muscle.
Why does a plasmid need an origin of replication?
The origin of replication also determines the plasmid’s compatibility: its ability to replicate in conjunction with another plasmid within the same bacterial cell. Plasmids that utilize the same replication system cannot co-exist in the same bacterial cell.
What are expression vectors PPT?
Expression Vectors: Vectors that can yield the protein products of the cloned genes. Two elements that are required for active gene expression: a strong promoter and a ribosome binding site near an initiating ATG codon.
How is plasmid replicated?
Plasmids utilize their host cell’s replication machinery in order to replicate. As described in our previous Origin of Replication post, DNA replication is initiated at the ORI and may be synchronized with the replication of the host cell’s chromosomal DNA or may be independent of the host’s cell cycle.
How many origins of replication do plasmids have?
Small organisms (e.g., bacteria), as well as plasmids and many viruses, have only one ori sequence per cellular genome (4.7 × 106 nucleotide pairs in E. coli), which is often an uninterrupted DNA molecule (Figs.
What is expression vector with example?
An example of expression vector is the plasmid used to produce insulin important for treating diseases such as diabetes. Another example is the expression vector that introduces specific genes necessary for the synthesis of beta-carotene into the cells of rice plants, giving rise to a new variety called golden rice.
What is T7 promoter sequence?
The T7 promoter is a sequence of DNA 18 base pairs long up to transcription start site at +1 (5′ – TAATACGACTCACTATAG – 3′) that is recognized by T7 RNA polymerase1 .
How is tac promoter formed?
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. It is commonly used for protein production in Escherichia coli. Two hybrid promoters functional in Escherichia coli were constructed.