Creation of genome-wide protein expression libraries using random activation of gene expression.

Here we report the use of random activation of gene expression to create genome-wide protein expression libraries. 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 duanyu1648 libraries created from multiple human cell lines, demonstrating that duanyu1648 libraries are inherently normalized. Pools of duanyu1648 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 duanyu1648 libraries and duanyu1648 expression clones are well suited for a number of biotechnological applications including gene discovery, protein characterization, drug development, and protein manufacturing.

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