Identification of genes expressed in human CD34+hematopoietic stem/progenitor cells by expressed sequence tags and efficient full-length cDNA cloning
| Autor: | Ya-Xin Wang, Shen Yu, Mao Mao, Zhu Chen, Li-Xin Kan, Qiu-Hua Huang, Ze-Guang Han, Gang Fu, Kai-Li He, Qinghua Zhang, Wu Jisheng, Yu Yaping, Jun Zhou, Sai-Juan Chen, Jian Gu, Bai-Wei Gu, Shu-Hua Xu |
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| Rok vydání: | 1998 |
| Předmět: |
Adenosine Triphosphatases
Genetics Regulation of gene expression Expressed sequence tag DNA Complementary Multidisciplinary Endosomal Sorting Complexes Required for Transport Molecular Sequence Data Vesicular Transport Proteins Antigens CD34 Biological Sciences Biology Hematopoietic Stem Cells Homology (biology) Repressor Proteins Gene Expression Regulation GenBank Complementary DNA Gene expression ATPases Associated with Diverse Cellular Activities Humans Genomic library Cloning Molecular Gene Gene Library |
| Zdroj: | Proceedings of the National Academy of Sciences. 95:8175-8180 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.95.14.8175 |
| Popis: | Hematopoietic stem/progenitor cells (HSPCs) possess the potentials of self-renewal, proliferation, and differentiation toward different lineages of blood cells. These cells not only play a primordial role in hematopoietic development but also have important clinical application. Characterization of the gene expression profile in CD34+HSPCs may lead to a better understanding of the regulation of normal and pathological hematopoiesis. In the present work, genes expressed in human umbilical cord blood CD34+cells were catalogued by partially sequencing a large amount of cDNA clones [or expressed sequence tags (ESTs)] and analyzing these sequences with the tools of bioinformatics. Among 9,866 ESTs thus obtained, 4,697 (47.6%) showed identity to known genes in the GenBank database, 2,603 (26.4%) matched to the ESTs previously deposited in a public domain database, 1,415 (14.3%) were previously undescribed ESTs, and the remaining 1,151 (11.7%) were mitochondrial DNA, ribosomal RNA, or repetitive (Alu or L1) sequences. Integration of ESTs of known genes generated a profile including 855 genes that could be divided into different categories according to their functions. Some (8.2%) of the genes in this profile were considered related to early hematopoiesis. The possible function of ESTs corresponding to so far unknown genes were approached by means of homology and functional motif searches. Moreover, attempts were made to generate libraries enriched for full-length cDNAs, to better explore the genes in HSPCs. Nearly 60% of the cDNA clones of mRNA under 2 kb in our libraries had 5′ ends upstream of the first ATG codon of the ORF. With this satisfactory result, we have developed an efficient working system that allowed fast sequencing of 32 full-length cDNAs, 16 of them being mapped to the chromosomes with radiation hybrid panels. This work may lay a basis for the further research on the molecular network of hematopoietic regulation. |
| Databáze: | OpenAIRE |
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