Expression of single chain antibodies (ScFvs) for c-myc oncoprotein in recombinant Escherichia coli membranes by using the ice-nucleation protein of Pseudomonas syringae.
The ice nucleation protein (INP) is a glycosyl phosphatidylinositol anchored outer membrane protein found in certain Gram-negative bacteria. In this study, the INP from Pseudomonas syringae was applied as a fusion partner with the single chain antibody fragment (ScFv) against the human oncoprotein c-myc. Two new plasmids pNinaZ-myc and pNinaZScFv-BsaA1 were constructed and cloned into Escherichia coli JM109. The expression of the fusion protein was successfully demonstrated in the cloned cells. The fusion proteins had no effect on the viability of the host cells. Ice nucleation activity measurements and flow cytometry studies were followed to investigate the membrane expression of the fusion protein.
Department of Chemical and Biochemical Engineering, Faculty of Engineering Science, The University of Western Ontario, London, Ontario, Canada N6A 5B9. abassi@julian.uwo.ca
A S
AS
Bassi
D N
DN
Ding
G B
GB
Gloor
A
A
Margaritis
D000595:Amino Acid Sequence; D000906:Antibodies / Q000235:genetics* / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002469:Cell Separation; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics*; D005434:Flow Cytometry; D008969:Molecular Sequence Data; D010957:Plasmids; D016271:Proto-Oncogene Proteins c-myc / Q000276:immunology*; D011549:Pseudomonas / Q000235:genetics*; D011995:Recombination, Genetic
D000906:Antibodies; D001425:Bacterial Outer Membrane Proteins; D016271:Proto-Oncogene Proteins c-myc; C063113:ice nucleation protein