| RRC ID |
33501
|
| 著者 |
Büssow K, Cahill D, Nietfeld W, Bancroft D, Scherzinger E, Lehrach H, Walter G.
|
| タイトル |
A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library.
|
| ジャーナル |
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.
|
| 巻・号 |
26(21)
|
| ページ |
5007-8
|
| 公開日 |
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
|
| 引用数 |
207
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 原核生物(大腸菌) |
pQE30NST
pQStrep2
pQStrep4
pQTEV
pSE111 |
