应用大规模测序技术和生物信息学研究造血干/祖细胞的基因表达及新基因的识别和克隆

Study on Gene Expression of Hematopoietic Stem/Progenitor Cells and Identification and Cloning of Novel Genes with Large-scale Sequencing and Bioinformatics

从新生儿脐血和成人骨髓中分选出造血干／祖细胞（ＨＳＣ／ＨＰＣ），构建成ｃＤＮＡ文库，对其进行大规模表达序列标签（ＥＳＴ）测序，通过生物信息学等手段分析基因表达谱，并进行新基因的全长ｃＤＮＡ克隆。在所测的１０５１２条可分析ＥＳＴ序列中，有９８６６条来自脐血ＣＤ３４＋细胞，其中４６９７条（４７６％）为已知基因，２６０３条（２６４％）为已知ＥＳＴ，１４１５条（１４３％）代表未知ＥＳＴ。在已知基因中，８２％基因与造血相关，２２７％涉及细胞代谢、结构和迁移，１３０％与细胞分裂和防御相关，２６２％与ＲＮＡ、蛋白质的合成相关，１０６％和细胞信号传递有关。对一些已知和未知的ＥＳＴ，综合测序、生物信息学等方法，进行全长克隆，已获得２３个新基因的全长ｃＤＮＡ。

Hematopoietic stem/progenitor cells were isolated from umbilical cord blood and adult bone marrow, and subject to cDNA library construction The gene expression pattern in CD34 + cells and the identification and cloning of novel genes were performed by sequencing ESTs and analyzing them with the tools of bioinformatics Among the obtained 10 512 ESTs which could be further analyzed, 9,866 were from umbilical cord blood where 4 697(67 6%) were known genes, 2 603(26 4%) were known ESTs and 1 415(14 3%) represented novel ESTs Within the identified genes, 8 2% was involved in hematopoiesis, 22 7% was associated with cell metabolism, structure and mobility, 13 0% was linked to cell division and defence, 26 2% was related to RNA protein synthesis and 10 6% was related with cell signal transduction In parallel, we developed an efficient working system combining sequencing, bioinformatics, etc and obtained 23 full-length cDNAs from both known and novel ESTs identified in this work