?

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