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- Title
A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library.
- Authors
Büssow, K; Cahill, D; Nietfeld, W; Bancroft, D; Scherzinger, E; Lehrach, H; Walter, G
- Abstract
We have developed a technique to establish catalogues of protein products of arrayed cDNA clones identified by DNA hybridisation or sequencing. A human fetal brain cDNA library was directionally cloned in a bacterial vector that allows IPTG-inducible expression of His6-tagged fusion proteins. Using robot technology, the library was arrayed in microtitre plates and gridded onto high-density in situ filters. A monoclonal antibody recognising the N-terminal RGSH6sequence of expressed proteins (RGS.His antibody, Qiagen) detected 20% of the library as putative expression clones. Two example genes, GAPDH and HSP90alpha, were identified on high-density filters using DNA probes and antibodies against their proteins.
- Publication
Nucleic acids research, 1998, Vol 26, Issue 21, p5007
- ISSN
0305-1048
- Publication type
Journal Article
- DOI
10.1093/nar/26.21.5007