|Título||Expression profiling using a hexamer-based universal microarray.|
|Publication Type||Journal Article|
|Year of Publication||2004|
|Autores||Roth, ME, Feng, L, McConnell, KJ, Schaffer, PJ, Guerra, CE, Affourtit, JP, Piper, KR, Guccione, L, Hariharan, J, Ford, MJ, Powell, SW, Krishnaswamy, H, Lane, J, Guccione, L, Intrieri, G, Merkel, JS, Perbost, C, Valerio, A, Zolla, B, Graham, CD, Hnath, J, Michaelson, C, Wang, R, Ying, B, Halling, C, Parman, CE, Raha, D, Orr, B, Jedrzkiewicz, B, Liao, J, Tevelev, A, Mattessich, MJ, Kranz, DM, Lacey, M, Kaufman, JC, Kim, J, Latimer, DR, Lizardi, PM|
|Date Published||2004 Apr|
|Palabras clave||3' Untranslated Regions, Algorithms, Animals, DNA Fragmentation, DNA Restriction Enzymes, DNA, Complementary, Galactose, Gene Expression Profiling, Humans, Image Processing, Computer-Assisted, Mice, Models, Genetic, Muscle, Skeletal, Muscles, Oligonucleotide Array Sequence Analysis, Promoter Regions, Genetic, Reverse Transcriptase Polymerase Chain Reaction, RNA, Messenger, Saccharomyces cerevisiae, Sequence Analysis, DNA, T-Lymphocytes, Transgenes|
We describe a transcriptional analysis platform consisting of a universal micro-array system (UMAS) combined with an enzymatic manipulation step that is capable of generating expression profiles from any organism without requiring a priori species-specific knowledge of transcript sequences. The transcriptome is converted to cDNA and processed with restriction endonucleases to generate low-complexity pools (approximately 80-120) of equal length DNA fragments. The resulting material is amplified and detected with the UMAS system, comprising all possible 4,096 (4(6)) DNA hexamers. Ligation to the arrays yields thousands of 14-mer sequence tags. The compendium of signals from all pools in the array-of-universal arrays comprises a full-transcriptome expression profile. The technology was validated by analysis of the galactose response of Saccharomyces cerevisiae, and the resulting profiles showed excellent agreement with the literature and real-time PCR assays. The technology was also used to demonstrate expression profiling from a hybrid organism in a proof-of-concept experiment where a T-cell receptor gene was expressed in yeast.
|Alternate Journal||Nat. Biotechnol.|