Decay-accelerating factor CD55 is identified as the receptor for echovirus 7 using CELICS, a rapid immuno-focal cloning method.
Using an anti-receptor mAb that blocks the attachment of echovirus 7 and related viruses (echoviruses 13, 21, 29 and 33), we have isolated a complementary DNA clone that encodes the human decay-accelerating factor (CD55). Mouse cells transfected with the CD55 clone bind echovirus 7, and this binding is blocked by the anti-receptor mAb. The method used (CELICS) allows rapid and direct cloning of genes encoding cell surface receptors. It is based on episomal replication and high efficiency expression of complementary DNA clones in the vector pCDM8 in COS or WOP cells, in conjunction with a sensitive immuno-focal screen that uses antibody probes linked to beta-galactosidase. Receptor positive cells were identified by a colour change and isolated individually using a micromanipulator. DNA extracted from a small number of cells was then cloned directly in Escherichia coli.
Ward
T
T
School of Animal and Microbial Sciences, University of Reading, Whiteknights, UK.
Pipkin
P A
PA
Clarkson
N A
NA
Stone
D M
DM
Minor
P D
PD
Almond
J W
JW
D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D018958:CD55 Antigens; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D029822:Enterovirus B, Human; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics*; D011991:Receptors, Virus / Q000235:genetics*; D012641:Selection, Genetic; D014162:Transfection
D015703:Antigens, CD; D018958:CD55 Antigens; D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D018799:Intercellular Adhesion Molecule-1