| 摘要: |
| 目的：克隆巨核白血病细胞诱导分化后表达变化的mRNA，从中寻找对诱导分化起关键作用的基因，最终揭示巨核白血病细胞诱导分化的分子机制。方法：人巨核白血病细胞HIMeg经13-顺式维甲酸处理4 d后，收集细胞总RNA，进行mRNA差别显示、cDNA片段克隆和序列分析。结果：获得5个差示cDNA片段，经RNA点杂交分析后发现其中3个为假阳性，1个因没有杂交信号而无法确定，只有1个cDNA片段得到证实，其对应的mRNA在诱导分化后表达下降。该cDNA片段的核苷酸序列已在GenBank中登录，登录号为AF026526。结论：克隆到一个未知基因的cDNA片段，其全长cDNA序列、基因结构及生物功能有待进一步研究。 |
| 关键词: 巨核细胞 白血病 分化 基因 克隆 |
| DOI： |
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| 基金项目: |
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| Cloning of differentially expressed genes in a human leukemic megakaryocytic cell line HIMeg after induced differentiation using mRNA differential display |
| 陆德如,程涛,童书鹏,李川晟,林文,戴建新,韩建中 |
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| Abstract: |
| Objective: In a human leukemic megakaryocytic cell line HIMeg, differentially expressed genes regulated by induced differentiation will be cloned, and from which the key genes in the process of induced differentiation will be identified. Methods: The human megakaryocytic leukemia cell line HIMeg was treated with 13-cis retinoic acid for 4 d, then total cellular RNA was extracted and reverse-transcribed to first strand cDNA. The first strand cDNA was subjected to PCR-based mRNA differential display, cloning and sequence analysis. Results: From 5 cDNA fragment candidates obtained, RNA dot blot analysis demonstrated non-regulation in 3 fragments, undetectable signals in one fragment, and altered gene expression in one cDNA fragment which was designated MDI-1 (mRNA downregulated after induced differentiation).The nucleotide sequence data reported here will appear in the GenBank under the accession number AF026526. Conclusion: A cDNA fragment encoded by a unknown gene was cloned. Further studies are required to determine its full length sequence, gene structure and biological function(s). |
| Key words: megakaryocyte leukemia differentiation gene cloning |
