gfp -gene as cDNA in a host-DNA-fragment E. coli (new host)
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Transcript of gfp -gene as cDNA in a host-DNA-fragment E. coli (new host)
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gfp-gene as cDNA in a host-DNA-fragment E. coli (new host)
gfp
?
???????
Aequorea victoria (donor)
host
gfp-gene as cDNA in a host-DNA-fragment DNA-transport system E.coli (new host)
gfp
host
gfp-gene as cDNA in a host-DNA-fragment DNA-transport system E.coli (new host)
1
3
cleavage of the interesting gene withappr. restriction enzymes
gfp
purified cloning vector DNApreparationof DNA-incor-porating cells
gfp
4
host
gfp-gene as cDNA in a host-DNA-fragment DNA-transport system E. coli (new host)
1
3
cleavage of the interesting gene withappr. restriction enzymes
gfp
purified cloning vector DNA preparationof DNA-incor-porating cells
gfp???
4
?
?
identification/verification???
host
gfp-gene as cDNA in a host-DNA-fragment DNA-transport system E. coli (new host)
1
3
cleavage of the interesting gene withappr. restriction enzymes
gfp
purified cloning vector DNA preparationof DNA-incor-porating cells(competent cells)
gfp
4
gfp
2
joining of vector DNAand gene fragment (ligation)
cleavage of vector DNA importation of
constructed DNA(transformation)
cultivation of cells
3
electrophoresis
recovery of gfp-fragment from agarose gel
gfp
gfp
?
host
4