Expression cloning of cDNA by phage display selection.
Expression cloning of a mouse kappa chain fragment has been achieved from a cDNA library by display of expressed proteins on filamentous phage and affinity selection for binding to anti-mouse Fab antibodies. Expressed proteins were anchored to the phage coat by a synthetic, anti-parallel leucine zipper, which had been selected from a semi-randomized zipper library for the ability to connect a test protein to phage. From a library of 4 x 10(6) transformants, two separate clones displaying different size cDNA inserts were recovered after four selection rounds. These results further demonstrate the utility of phage display for cDNA expression cloning.
Light
J
J
The Burnham Institute, La Jolla, CA 92037, USA. light@ljcrf.edu
Maki
R
R
Assa-Munt
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D016350:Leucine Zippers / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library
D018076:DNA, Complementary; D007140:Immunoglobulin Fab Fragments; D007145:Immunoglobulin kappa-Chains; D019151:Peptide Library