| Hematopoiesis is a complicated process which is directed accurately by many factors. Interference with the key factors can lead to loss of lineage identity, increased infidelity, lineage reprogramming and even malignant transformation. In this work, we sequenced whole transcriptome using the new generation sequencing technology (SOLiD) to perform comparative analysis of gene expression profiling among three myeloid cell lines(K562,HL-60and THP1) to investigate their transcriptome characteristic, functional formation mechanism and differences between the acute and chronic myeloid leukemia. K562was obtained from a chronic myeloid leukemia patient representing early stage of erythroid; HL-60was obtained from an acute promyelocytic leukemia patient representing promyelocyte; THP1was obtained from an acute monocytic leukemia patient representing monocyte.Here, we reported37183840,21910289and23594340mapped50bp reads separately from the three samples. In all these three libraries14120740(51.56%),8561613(52.53%),9488637(51.70%) were found in the exonic regions. Based on the selection (tag number>5in exonic regions), we identified16779,14028and13448genes that are expressed in the three libraries. We used RPKM values to calculate the gene expression abundance, more than70%of which are between1and50in these three libraries. High-expressed genes are enriched in translation; medium-expressed genes are those for basal metabolism; and low-expressed genes are involved in development and regulation.Differential expressed gene analysis suggested that genes expressed in K562are enriched for the maintenance of its self-renewal capacity, but there are also some genes representative of erythroid cells; while HL-60and THP1have more genes expressed for their immune function. There some differences in the performance between chronic and acute myeloid leukemia. In chronic myeloid leukemia, the up-regulated pathway may lead to proliferation; while differentiation block in acute myeloid leukemia. The microsatellite enriched library of small abalone (Haliotis diversicolor Reeve) was constructed according to the strong affinity between biotin and streptavidin and genomic DNA was fragmented with ultrasonication, which is which is according Li et al. Linker-ligated DNA fragments were hybridized with biotinylated microsatellite probes, and then subjected to streptavidin-coated magnetic beads. PCR amplification was performed to obtain double-stranded DNA fragments containing microsatellite. Ligation and transformation were carried out by using the pGEM-T Vector SystemⅠ and Escherichia coli DH10B competent cell.480positive clones were selected for bidirectional sequencing and504contigs were assembled.418contigs contained microsatellite site, accounting for82.93%which indicat a high degree of microsatellite enrichment。 We used Primer3.0to design primers and amplify5individuals from different geographic populations.2%agarose gel was used to determine annealing temperature. Finally, we acquired170pair of primers can be used. Eleven novel microsatellite markers were tested on22individuals from two different geographic populations. We identified a total of162alleles from the11microsatellite loci. All of the loci were highly polymorphic. Polymorphism information content (PIC) is ranging from0.7276to0.9163. Observed and expected heterozygosities ranged from0.2727to1.0000and from0.7738to0.9429, respectively. Three loci deviated significantly from Hardy-Weinberg equilibrium. No pairs of loci displayed linkage disequilibrium. These polymorphic markers will be used to analyze population structure, genetic diversity and construct a genetic linkage map. |
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