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A selectable marker is a gene introduced into cells, especially bacteria or cells in culture, which confers one or more traits suitable for artificial selection.They are a type of reporter gene used in laboratory microbiology, molecular biology, and genetic engineering to indicate the success of a transfection or transformation or other procedure meant to introduce foreign DNA into a cell.
In bacteria, antibiotics are used almost exclusively. In plants, antibiotics that kill the chloroplast are often used as well, although tolerance to salts and growth-inhibiting hormones is becoming more popular. In mammals, resistance to antibiotics that would kill the mitochondria is used as a selectable marker.
This enzyme causes bacteria expressing the gene to appear blue when grown on a medium that contains the substrate analog X-gal. An example of a selectable marker which is also a reporter in bacteria is the chloramphenicol acetyltransferase (CAT) gene, which confers resistance to the antibiotic chloramphenicol. [11]
Some vectors contain two selectable markers, for example the plasmid pACYC177 has both ampicillin and kanamycin resistance gene. [6] Shuttle vector which is designed to be maintained in two different organisms may also require two selectable markers, although some selectable markers such as resistance to zeocin and hygromycin B are effective in ...
Ligation of selectable marker into plasmid vector: this allows for the differential selection of colonies with, or without the marker gene. An antibiotic resistance gene allows the YAC vector to be amplified and selected for in E. coli by enabling E. coli containing the YAC vector to survive in the presence of an antibiotic.
When bacterial cells are used as host organisms, the selectable marker is usually a gene that confers resistance to an antibiotic that would otherwise kill the cells, typically ampicillin. Cells harboring the plasmid will survive when exposed to the antibiotic, while those that have failed to take up plasmid sequences will die.
The T-DNA portion of the binary vector is flanked by left and right border sequences and may include a transgene as well as a plant selectable marker. Outside of the T-DNA, the binary vector also contains a bacterial selectable marker and an origin of replication (ori) for bacteria. [4] Representative series of binary vectors are listed below.
Since URA3 allows for both positive and negative selection, it has been developed as a genetic marker for DNA transformations and other genetic techniques in bacteria and many fungal species. It is one of the most important genetic markers in yeast genetic modification. While URA3 is a powerful selectable marker, it has a high background.
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