A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library. |
| |
| Authors: | K Bü ssow, D Cahill, W Nietfeld, D Bancroft, E Scherzinger, H Lehrach, G Walter |
| |
| Affiliation: | K Büssow, D Cahill, W Nietfeld, D Bancroft, E Scherzinger, H Lehrach, and G Walter |
| |
| 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. |
| |
| Keywords: | |
|
|
