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Auxotrophic genetic markers are often used in molecular genetics; they were famously used in Beadle and Tatum's Nobel prize-winning work on the one gene-one enzyme hypothesis, connecting mutations of genes to protein mutations. This then allows for biosynthetic or biochemical pathway mapping that can help determine which enzyme or enzymes are ...
Negative selection through replica plating to screen for ampicillin sensitive colonies. Replica plating is a microbiological technique in which one or more secondary Petri plates containing different solid (agar-based) selective growth media (lacking nutrients or containing chemical growth inhibitors such as antibiotics) are inoculated with the same colonies of microorganisms from a primary ...
URA3 is often used in yeast research as a "marker gene", that is, a gene to label chromosomes or plasmids. URA3 encodes Orotidine 5'-phosphate decarboxylase (ODCase), which is an enzyme that catalyzes one reaction in the synthesis of pyrimidine ribonucleotides (a component of RNA). [1]
Auxotrophic selection markers that allow an auxotrophic organism to grow in minimal growth medium may also be used; examples of these are LEU2 and URA3 which are used with their corresponding auxotrophic strains of yeast. [7] Another kind of selectable marker allows for the positive selection of plasmid with cloned gene.
Selectable markers allow scientists to separate non-recombinant organisms (those which do not contain the selectable marker) from recombinant organisms (those which do); that is, a recombinant DNA molecule such as a plasmid expression vector is introduced into bacterial cells, and some bacteria are successfully transformed while some remain non-transformed.
The pyrE − mutant is an auxotrophic mutant for uracil. The pyrE from a distantly related genus of T. maritima rescued the uracil auxotrophy of the pyrE − mutant of T. maritima and has been proven to be a suitable marker. For the first time, the use of this marker allowed the development of an arabinose (araA) mutant of T. maritima.
Next to the above-mentioned selection makers a few auxotrophic strains were generated to work with auxotrophic makers. The URA3 marker (URA3 blaster method) is an often-used strategy in uridine auxotrophic strains; however, studies have shown that differences in URA3 position in the genome can be involved in the pathogeny of C. albicans. [119]
All in all, homologous recombination provides more specificity when creating a mutant strain. Depending on the mutant, auxotrophy markers (requires lost gene to be inserted) or prototrophy (when causing essential gene deletion) be used for selection.