258
FRIEDHELM HILDEBRANDT
PROTEIN
..
POST-PURI FICATION
CLONING
The protein has been
identified (pu rified)
CLONING STRATEGIES
GENE
CLONING BY
HOMOLOGY
. .
Sequence similarity
with a known gene
Antibodies
I
~ ~,~,.~~
screening of:
,.-~-0 -1-ig_o_n_u_c_le_o_b_.d_e_s--,
- prokaryotic
expression
I i brary (l.gt I I )
- eukaryo tic
expression
library (co cells)
'
- screeni ng of
cDNA libraries
- PCR
FUNCTION
CLONING BY
EXPRESSION
Presence of the function
can be easily detected in
eukaryotic cells
t
the function is not present
in the recipient cell
t
transfection
mRNA
OTHER STRATEGIES
genomic DNA
cDNA
POSITIONAL
CLONING
DIFFERENTIAL
CLONING
YEAST 2-HYBRID SYSTEM
(protei n interactions in yeast)
t
t
ldentilication of transfectants
t
t
isolation of
"sib" selection
transfectants
screenin g of a
genomic library
Fig. I. Summary of cloning strategies (modified after C. Sardet, In: Molecular Medicine, Freiburg, 1994)