Linear expression elements: a rapid, in vivo, method to screen for gene functions.
The increasing accumulation of genomic sequence information has accentuated the need for new methods to efficiently assess gene function and to prepare reagents to study these functions. Toward solving this general problem in functional genomics, we report a method by which any PCR-amplified open-reading frame (ORF) can be noncovalently linked to a eukaryotic promoter and terminator, and directly injected into animals to produce local gene expression. We also demonstrate that ORFs can be delivered into mice to produce antibodies specific for the encoded foreign protein by simply attaching mammalian promoter and terminator sequences. This technology makes it possible to screen large numbers of genes rapidly for their functions in vivo or to produce immune responses without the necessity of cloning, bacterial propagation, or protein purification.
Sykes
K F
KF
Department of Internal Medicine, University Texas-Southwestern Medical Center, Dallas 75235-8573, USA.
Johnston
S A
SA
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000276:immunology; D000941:Antigens / Q000235:genetics; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D051379:Mice; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology; D016366:Open Reading Frames / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D013728:Terminator Regions, Genetic; D019444:Vaccines, DNA / Q000276:immunology
D000906:Antibodies; D000907:Antibodies, Bacterial; D000941:Antigens; D000942:Antigens, Bacterial; D019444:Vaccines, DNA