دورية أكاديمية
Recovery of developmentally defined gene sets from high-density cDNA macroarrays.
| العنوان: | Recovery of developmentally defined gene sets from high-density cDNA macroarrays. |
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| المؤلفون: | Rast JP; Division of Biology 156-29, California Institute of Technology, Pasadena, California 91125, USA., Amore G, Calestani C, Livi CB, Ransick A, Davidson EH |
| المصدر: | Developmental biology [Dev Biol] 2000 Dec 15; Vol. 228 (2), pp. 270-86. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: United States NLM ID: 0372762 Publication Model: Print Cited Medium: Print ISSN: 0012-1606 (Print) Linking ISSN: 00121606 NLM ISO Abbreviation: Dev Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: San Diego, CA : Elsevier Original Publication: New York. |
| مواضيع طبية MeSH: | Gene Expression Regulation, Developmental* , Gene Library* , Oligonucleotide Array Sequence Analysis*, Embryo, Nonmammalian/*physiology , Sea Urchins/*embryology , Sea Urchins/*genetics, Animals ; Animals, Genetically Modified ; DNA Probes ; DNA, Complementary ; Genes, Reporter ; Green Fluorescent Proteins ; Luminescent Proteins/analysis ; Luminescent Proteins/genetics ; RNA, Messenger/genetics ; Transcription Factors/metabolism |
| مستخلص: | New technologies for isolating differentially expressed genes from large arrayed cDNA libraries are reported. These methods can be used to identify genes that lie downstream of developmentally important transcription factors and genes that are expressed in specific tissues, processes, or stages of embryonic development. Though developed for the study of gene expression during the early embryogenesis of the sea urchin Strongylocentrotus purpuratus, these technologies can be applied generally. Hybridization parameters were determined for the reaction of complex cDNA probes to cDNA libraries carried on six nylon filters, each containing duplicate spots from 18,432 bacterial clones (macroarrays). These libraries are of sufficient size to include nearly all genes expressed in the embryo. The screening strategy we have devised is designed to overcome inherent sensitivity limitations of macroarray hybridization and thus to isolate differentially expressed genes that are represented only by low-prevalence mRNAs. To this end, we have developed improved methods for the amplification of cDNA from small amounts of tissue (as little as approximately 300 sea urchin embryos, or 2 x 10(5) cells, or about 10 ng of mRNA) and for the differential enhancement of probe sequence concentration by subtractive hybridization. Quantitative analysis of macroarray hybridization shows that these probes now suffice for detection of differentially expressed mRNAs down to a level below five molecules per average embryo cell. (Copyright 2000 Academic Press.) |
| معلومات مُعتمدة: | GM 18478 United States GM NIGMS NIH HHS; HD-37105 United States HD NICHD NIH HHS |
| المشرفين على المادة: | 0 (DNA Probes) 0 (DNA, Complementary) 0 (Luminescent Proteins) 0 (RNA, Messenger) 0 (Transcription Factors) 147336-22-9 (Green Fluorescent Proteins) |
| تواريخ الأحداث: | Date Created: 20001209 Date Completed: 20010202 Latest Revision: 20071114 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1006/dbio.2000.9941 |
| PMID: | 11112329 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 0012-1606 |
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| DOI: | 10.1006/dbio.2000.9941 |