Cloning and functional analysis of cDNAs with open reading frames for 300 previously undefined genes expressed in CD34+hematopoietic stem/progenitor cells

Three hundred cDNAs containing putatively entire open reading Frames (ORFs) For previously undefined genes were obtained From CD34+ hematopoietic stem /progenitor cells (HSPCs), based on EST cataloging, clone sequencing, in si lico cloning, and rapid amplification of cDNA ends (RACE). The cDNA sizes r anged from 360 to 3496 bp and their ORFs coded for peptides of 58-752 amino acids. Public database search indicated that 225 cDNAs exhibited sequence similarities to genes identified across a variety of species. Homology anal ysis led to the recognition of 50 basic structural motifs/domains among the se cDNAs. Genomic exon-intron organization could be established in 243 gent s by integration of cDNA data with genome sequence information. Interesting ly, a new gene named as HSPC070 on 3p was found to share a sequence of 105b p in 3' UTR with RAF gene in reversed transcription orientation. Chromosoma l localizations were obtained using electronic mapping for 192 genes and wi th radiation hybrid (RH) for 38 genes. Macroarray technique was applied to screen the gene expression patterns in five hematopoietic cell lines (NB4 H L60, U937, K562, and Jurkat) and a number of genes with differential expres sion were found. The resource work has provided a wide range of information useful not only For expression genomics and annotation of genomic DNA sequ ence, but also For further research on the function of genes involved in he matopoietic development and differentiation.