|
Authors
|
 |
J.J.Harrington,
B.Sherf,
S.Rundlett,
P.D.Jackson,
R.Perry,
S.Cain,
C.Leventhal,
M.Thornton,
R.Ramachandran,
J.Whittington,
L.Lerner,
D.Costanzo,
K.McElligott,
S.Boozer,
R.Mays,
E.Smith,
N.Veloso,
A.Klika,
J.Hess,
K.Cothren,
K.Lo,
J.Offenbacher,
J.Danzig,
M.Ducar.
|
|
Here we report the use of random activation of gene expression (RAGE) to create
genome-wide protein expression libraries. RAGE libraries containing only 5 x
10(6) individual clones were found to express every gene tested, including genes
that are normally silent in the parent cell line. Furthermore, endogenous genes
were activated at similar frequencies and expressed at similar levels within
RAGE libraries created from multiple human cell lines, demonstrating that RAGE
libraries are inherently normalized. Pools of RAGE clones were used to isolate
19,547 human gene clusters, approximately 53% of which were novel when tested
against public databases of expressed sequence tag (EST) and complementary DNA
(cDNA). Isolation of individual clones confirmed that the activated endogenous
genes can be expressed at high levels to produce biologically active proteins.
The properties of RAGE libraries and RAGE expression clones are well suited for
a number of biotechnological applications including gene discovery, protein
characterization, drug development, and protein manufacturing.
|