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

Konrad Büssow | Zendy

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

Author(s) -

Konrad Büssow

Publication year - 1998

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/26.21.5007

Subject(s) - biology , complementary dna , cdna library , microbiology and biotechnology , fusion protein , genomic library , monoclonal antibody , lac operon , gene , antibody , gene expression , recombinant dna , genetics , peptide sequence

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.

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