RRC ID 33501
Author Büssow K, Cahill D, Nietfeld W, Bancroft D, Scherzinger E, Lehrach H, Walter G.
Title A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library.
Journal Nucleic Acids Res
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.
Volume 26(21)
Pages 5007-8
Published 1998-11-1
DOI 10.1093/nar/26.21.5007
PII gkb763
PMID 9776767
PMC PMC147919
MeSH Antibodies, Monoclonal* Brain Chemistry Cloning, Molecular DNA, Complementary / genetics* Fetus Gene Expression Gene Library* Humans Molecular Sequence Data Nerve Tissue Proteins / genetics Nerve Tissue Proteins / immunology Proteins / genetics* Proteins / immunology* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / immunology
IF 11.502
Times Cited 207
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
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Prokaryotes E. coli pQE30NST pQStrep2 pQStrep4 pQTEV pSE111