. While perusing the E. coli K12 genome sequence,you come across a gene with no known function. Theamino acid sequence of the gene’s protein productshows weak similarities with known porins, proteinsthat cross a cellular membrane to let molecules suchas amino acid or sugar nutrients (or drugs like penicillin) pass through. Some porins are nonspecific and letany solute up to a certain size transit into the cell.Other porins are specific and allow the transit ofcertain sugars but not others. What genetic experiments could you do to try to determine whether thisnew gene has a specific function in allowing bacterialcells to scavenge the sugar maltose from the environment? Describe scenarios that might complicate yourexperimental approach.
. While perusing the E. coli K12 genome sequence,
you come across a gene with no known function. The
amino acid sequence of the gene’s protein product
shows weak similarities with known porins, proteins
that cross a cellular membrane to let molecules such
as amino acid or sugar nutrients (or drugs like penicillin) pass through. Some porins are nonspecific and let
any solute up to a certain size transit into the cell.
Other porins are specific and allow the transit of
certain sugars but not others. What genetic experiments could you do to try to determine whether this
new gene has a specific function in allowing bacterial
cells to scavenge the sugar maltose from the environment? Describe scenarios that might complicate your
experimental approach.
Genome is defined as an organism’s complete set of DNA with all of its genes present. This will contain all the needed information for the organism to survive and grow.
We could try to generate a null mutation in the gene by gene targeting technique which uses homologous recombination in order to modify an endogenous gene.
Using recombinant DNA technology, a DNA construct is being created in which a drug resistance gene is flanked on one side by sequences at the 5′ end of the gene in question, and on the other side by sequences from the 3′ end of the gene. Then these linear DNA fragments are introduced into wild-type E. coli. The selection of colonies containing the drug resistance was done by allowing the cells to grow on agar media containing the drug in petri plates. Colonies which appear on the plates have a null mutation in the gene which get inherited from a bacterium that incorporated, by the process of homologous recombination.
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