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为了进一步研究长期培养体系 (long termculture ,LTC)中造血基质细胞调控造血的分子基础及其作用机制 ,并发现新型造血调控因子 ,以LTC体系培养后小鼠骨髓造血基质细胞 (LTC St)为被扣除杂交组 ,以未经培养的小鼠骨髓细胞 (BMC)为扣除杂交组 ,采用抑制性扣除杂交法 (SSH)进行杂交 ,将杂交产物克隆并测序 ,进行生物信息学分析。结果 :获得了 131个差异表达EST克隆 ,这些克隆代表了 2 6种已知或部分已知的不同基因和 7条无任何线索的全新基因 ,5条具有完整开放阅读框架 ,无功能线索的新基因中 3条有信号肽。结论 :通过本实验得到了LTC体系中特异性表达的一个消减文库 ,获得了几条可能与造血调控相关的新型基因或EST ,为揭示造血微环境支持造血的分子机制提供了有意义的线索
In order to further study the molecular basis and mechanism of hematopoietic stem cell hematopoiesis in long term culture (LTC) and to find novel hematopoietic regulatory factors, mouse bone marrow hematopoietic stromal cells (LTC St) cultured in LTC system After subtracting the hybridization group, the uncultured mouse bone marrow cells (BMC) were subtracted from the hybridization group, hybridization was performed by SSH, and the hybridization products were cloned and sequenced for bioinformatics analysis. RESULTS: A total of 131 differentially expressed EST clones were obtained. These clones represented 26 novel genes known or partially known and 7 novel genes without any clue. Five novel genes with a complete open reading frame and no functional cues Three genes in the signal peptide. CONCLUSIONS: A subtractive library specifically expressing in LTC system was obtained in this experiment, and several new genes or ESTs that may be related to hematopoietic regulation were obtained, which provided a meaningful clues for revealing the molecular mechanism of hematopoietic microenvironment supporting hematopoiesis