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Expression of Single Chain Antibodies (ScFvs) for c‐myc Oncoprotein in Recombinant Escherichia coli Membranes by Using the Ice‐Nucleation Protein of Pseudomonas syringae
Author(s) -
Bassi Amarjeet S.,
Ding Daniel N.,
Gloor Gregory B.,
Margaritis Argyrios
Publication year - 2000
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp000053k
Subject(s) - pseudomonas syringae , escherichia coli , fusion protein , recombinant dna , microbiology and biotechnology , chemistry , plasmid , flow cytometry , bacterial outer membrane , biology , bacteria , biochemistry , gene , genetics
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‐ Bsa A1 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.