Cloning And Function Prediction Of The Full-length CDNA Of A Novel Gene (FAMLF) Identified In Familial Acute Myelogenous Leukemia

Acute leukemia is a common malignant tumour of hematological system. Generation and development of leukemia are closely associated with abnormality of structure and function of the related gene, cloning of novel leukemia related gene can help to elucidate molecular biological mechanisms of leukemogenesis ,it also provide a base for developing diagnostic agents and clinical therapies of leukemia.【Objective】To clone the full-length cDNA of a novel gene of familial acute myelogenous leukemia and to explain the molecular mechanisms of the disease at the gene level.【Method】our laboratory constructed a subtractive cDNA pool of familial acute myelogenous leukemia using the method of suppression subtractive hybridization (SSH) and obtained 28 positive clones; 11 were verified novel unknown EST by NCBI homology analysis .We select a novel unknown differential EST (GenBank accession number :CV973101), the experiment was designed as follow:1. detect differential expression in familial acute myelogenous leukemia patient sample and health adult sample with one-step semi-quantitative reverse transcriptase-PCR (RT-PCR);2. clone the full-length cDNA by SMART-rapid amplification of cDNA ends(SMART-RACE);3.predict novel gene associated function by bioinformatics strategy.【Results】Full-length cDNA of 2313 bp was obtained with complete open reading frame(ORF) of 249bp ,it was a novel gene whose function was unknown by NCBI BLAST ,localized on 1q31.3 of human chromosome and coded a 82-amino acid polypeptide; novel gene was named Homo sapiens familial acute myelogenous leukemia related factor(FAMLF) and submitted to GenBank with the accession number: (Nucleotide)DQ359746 and (Protein) ABD15131.【Conclusion】(1) The full-length cDNA of a novel familial acute myelogenous leukemia associated gene (FAMLF)was acquired,it coded a 82-amino acid polypeptide and localized on 1q31.3 . (2) FAMLF is a novel gene whose function is unknown. (3) RACE is an effective approach to clone full-length cDNA. (4) Function of novel gene FAMLF can be predicted by bioinformatics.