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

被引：210

作者：

Büssow, K ^{[1
]}

Cahill, D ^{[1
]}

Nietfeld, W ^{[1
]}

Bancroft, D ^{[1
]}

Scherzinger, E ^{[1
]}

Lehrach, H ^{[1
]}

Walter, G ^{[1
]}

机构：

[1] Max Planck Inst Mol Genet, D-14195 Berlin, Germany

来源：

NUCLEIC ACIDS RESEARCH | 1998年 / 26卷 / 21期

关键词：

D O I：

10.1093/nar/26.21.5007

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 His(6)-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 RGSH(6) sequence of expressed proteins (RGS.His antibody, Qiagen) detected 20% of the library as putative expression clones. Two example genes, GAPDH and HSP90 alpha, were identified on high-density filters using DNA probes and antibodies against their proteins.

引用

页码：5007 / 5008

页数：2

共 7 条

[1] HIGH-LEVEL EXPRESSION OF RECOMBINANT GENES IN ESCHERICHIA-COLI IS DEPENDENT ON THE AVAILABILITY OF THE DNAY GENE-PRODUCT [J].