The Screen Of A Familial Acute Myelogenous Leukemia Relative Novel Gene With SNP Array, The Expression And The Construction Of Eukaryotic Expression System Of The Familial Acute Myelogenous Leukemia Relative Gene(FAMLF)

Acute leukemia is a common hematological malignancy. Generation and development of leukemia are closely associated with abnormality of structure and function of the related gene. This project is to define the novel leukemia associated genes of leukemia-high density family and to provide scientific proof for diagnosis and therapy for leukemia patients in a gene level. This research is contributed to the exploration of the pathogenesy of leukemia, the drug development for leukemia, and the improvement in the recovery rate of leukemia.【Objective】1.To screen and clone the familial acute myelogenous leukemia pathopoiesis relative genes and to explain the molecular mechanisms of the disease at the gene level. 2.To construction eukaryotic expression system base on our previous research for the functional study in future.【Method】1. With a positional cloning strategy,we performed the genetype analysis for the leukemic patients, high-risk individuals and healthy siblings from the familiar leukemia by using advanced 500k SNP microarrays. 2.A bioinformatics-based analysis with the associated statistical genetic software was applied to carry out linkage, haplotype, and copy number analysis to identify the chromosomal translocations link to familiar leukemia. 3.The candidate genes located in the target chromatin domain were identified. 4.The novel leukemia-associated gene in a chromatin domain with copy munbers change were identified integration of RT-PCR data. 5. Differential expression in familial acute myelogenous leukemia patient sample and health control were detect with one-step semi-quantitative reverse transcriptase-PCR (RT-PCR). 6.With engineered technique, the eukaryotic expression of FAMLF was constructed.【Results】1.Eight family members'genetyping of whole genome was obtained. 2.Through the linkage analysis and haplotype mapping, 21 genes with LOD higer than 1.5 and same sharing haplotype were found. 3.After copy number analysis, 56 genes with copy number change were defined. 4.RT-PCR expresson analysis confirmed with copy number analysis. 5.FAMLF was expressed high in 23 patients compared to 77 healthy control, and the expression level shows negative skewness distribution in healthy populations. 6. The eukaryotic expression system of FAMLF was constructed successfully.【Conclusion】1.Genes of linkage LOD more than 1.5 with sharing haplotype were acquired. 2. Genes with copy number change were obtained. 3.SNP array technology is one of the effective approach to position Mendelian inheritance genes. 4. Copy numbers analysis of SNP can be used to find genes with copy number change. 5.The expression of FAMLF is high in partly leukemic patients compared to that of healthy controls. 6. FAMLF eucaryon expression system was constructed successfully.