Engine - Model hEX1 -Protein Array

The hEX1 protein expression library was constructed by preparing cDNA from human fetal brain poly(A)+ RNA, using oligo (dT)-priming. The resulting fragments (larger than 500 bp) were cloned into a modified expression vector (pQE30NST) allowing protein expression to be induced by addition of IPTG to the growth medium. The resulting expressed proteins include an N-terminal RGS-HIS-Tag, which can be used for detection and purification purposes. 193.536 clones were picked, spotted and induced on PVDF-membranes and thereafter assayed for protein expression using an anti-HIS-antibody. Based on this screen, 37.830 putative expression clones were selected and re-arrayed into 384 well microtiter plates which constitute the hEX1-library. Spotted on high-density protein filters on two PVDF-membranes, the library is available as hEX1 part 1 and part 2 (Büssow et al., 1998).