marker genes Flashcards
Marker Genes
these are used to identify if a gene has been taken up by a bacterial cell
what do they involve
a second separate gene on the plasmid
what genes are included
those reistant to antibiotics, fluorescent protein , or produce an enzyme that is easily identifiable
what is cut first
the gene for resistance to antibiotic a
what is inserted into this gene
the dna from the cell that manufacture the desired gene
what will have to be the same
the same restriction endonucleases to create same sticky ends
what does the plasmid have now
a non functioning antibitotic resistance to a incorporation of the new gene
how do we identify the plasmids that have taken up the inserted gene
we use replic plating
how does the process work
the antibiotic resistant gene which produces an enzyme will no longer function
what do we grow them on
agar that contains the corresponding bacteria, those plasmids that don’t survive we know that they contain the desired genes
what do we then do
intentify the colonies that no longer have resistance to the antibiotic and replicate them
how do we use fluorescent markers
we transfer a gene from a jelly fish into the plasmid
what gene is responsible
the gene gfp produce a green fluorescent protein
how is the desired gene then implanted
the desired gene is cut using the same restriction enzymes used to cut the centre of the gfp and is inserted into the centre of this gene
how can we identify plasmids that has taken up desired gene
they will no longer fluoresce however those that have not taken up the gene will continue to fluoresce
