Lee S, Zhou G, Clark T, Chen J, Rowley J D, Wang S M
Department of Medicine, University of Chicago Medical Center, 5841 South Maryland, MC2115, Chicago, IL 60637, USA.
Proc Natl Acad Sci U S A. 2001 Mar 13;98(6):3340-5. doi: 10.1073/pnas.051013798.
We performed a genome-wide analysis of gene expression in primary human CD15(+) myeloid progenitor cells. By using the serial analysis of gene expression (SAGE) technique, we obtained quantitative information for the expression of 37,519 unique SAGE-tag sequences. Of these unique tags, (i) 25% were detected at high and intermediate levels, whereas 75% were present as single copies, (ii) 53% of the tags matched known expressed sequences, 34% of which were matched to more than one known expressed sequence, and (iii) 47% of the tags had no matches and represent potentially novel genes. The correct genes were confirmed by application of the generation of longer cDNA fragments from SAGE tags for gene identification (GLGI) technique for high-copy tags with multiple matches. A set of genes known to be important in myeloid differentiation were expressed at various levels and used different spliced forms. This study provides a normal baseline for comparison of gene expression in myeloid diseases. The strategy of using SAGE and GLGI techniques in this study has broad applications to the genome-wide identification of expressed genes.
我们对原代人CD15(+)髓系祖细胞进行了全基因组基因表达分析。通过使用基因表达序列分析（SAGE）技术，我们获得了37,519个独特SAGE标签序列表达的定量信息。在这些独特标签中，（i）25%在高水平和中等水平被检测到，而75%以单拷贝形式存在，（ii）53%的标签与已知表达序列匹配，其中34%与多个已知表达序列匹配，（iii）47%的标签无匹配，代表潜在的新基因。通过应用从SAGE标签生成更长的cDNA片段以进行基因鉴定（GLGI）技术对具有多个匹配的高拷贝标签进行分析，确认了正确的基因。一组已知在髓系分化中重要的基因以不同水平表达并使用不同的剪接形式。本研究为髓系疾病中基因表达的比较提供了正常基线。本研究中使用SAGE和GLGI技术的策略在全基因组范围内鉴定表达基因方面具有广泛应用。
