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Nucleic Acids Res. 1998 Nov 1;26(21):5007-8.

A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library.

Author information

  • 1Max Planck Institute for Molecular Genetics, Ihnestr. 73, D-14195 Berlin, Germany. buessow@mpimg-berlin-dahlem.mpg.de

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.

PMID:
9776767
[PubMed - indexed for MEDLINE]
PMCID:
PMC147919
Free PMC Article
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