Bacterial vectors to target and/or purify polypeptides: their use in immunological studies.
The construction of recombinant proteins by genetic engineering has opened new avenues in basic research (studies on protein organization, protein folding, immunogenicity of proteins, ...) and many different applications. Recombinant proteins which keep properties of both parental proteins are especially interesting. For example, if one protein--the vector protein--is targeted to a given cellular compartment, the other protein--the passenger--may be identically targeted. Also, if the vector protein can be purified by a simple affinity chromatographic procedure, this property may be extended to the passenger. The authors have developed a genetic procedure to detect "permissive" sites within potential vector proteins so that genetic fusion to these sites keep most or all biological properties of the vector. When they used LamB, an outer membrane protein from E. coli, foreign sequences could be expressed at the bacterial cell surface. This may lead to several types of applications: live bacterial vaccines, simple diagnostic tests, selection procedures for peptides with biological activity. When they used the MalE protein, a periplasmic maltose binding protein from E. coli, the passengers could be exported and purified in one-step high affinity chromatography in mild non-denaturing conditions. This led us to a simple preparation and purification scheme for the soluble part of the CD4 receptor for the Human Immunodeficiency Virus (HIV).
Clement
J M
JM
Unité de Programmation Moléculaire et Toxicologie Génétique (CNRS URA 271), Institut Pasteur, Paris.
Charbit
A
A
Martineau
P
P
O'Callaghan
D
D
Szmelcman
S
S
Leclerc
C
C
Hofnung
M
M
D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology; D015704:CD4 Antigens / Q000276:immunology / Q000302:isolation & purification; D004926:Escherichia coli / Q000276:immunology; D005822:Genetic Vectors; D015488:HIV Antigens / Q000276:immunology / Q000302:isolation & purification; D007158:Immunologic Techniques; D010455:Peptides / Q000302:isolation & purification*
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D015704:CD4 Antigens; D015488:HIV Antigens; D010455:Peptides