DECAY-ACCELERATING FACTOR CD55 IS IDENTIFIED AS THE RECEPTOR FOR ECHOVIRUS-7 USING CELICS, A RAPID IMMUNE-FOCAL CLONING METHOD

被引:137
作者
WARD, T
PIPKIN, PA
CLARKSON, NA
STONE, DM
MINOR, PD
ALMOND, JW
机构
[1] UNIV READING,SCH ANIM & MICROBIAL SCI,READING RG6 2AJ,BERKS,ENGLAND
[2] NATL INST BIOL STAND & CONTROLS,POTTERS BAR EN6 3QG,HERTS,ENGLAND
关键词
CD55; CELICS; CLONING METHOD; DAF; VIRUS RECEPTOR;
D O I
10.1002/j.1460-2075.1994.tb06836.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 immune-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.
引用
收藏
页码:5070 / 5074
页数:5
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