[Identification of differentially expressed genes in familial acute myelogenous leukemia by suppression subtractive hybridization].

OBJECTIVE

To isolate genes expressed differentially from the bone marrow cells of patients with familial acute myelocytic leukemia and to explain the molecular mechanisms of the disease at the gene level.

METHODS

Bone marrow cells were obtained from a family with cases of familial acute myelocytic leukemia for successive 4 generations. Super SMART cDNA synthesis and suppression subtractive hybridization (SSH) were performed to set up a subtractive cDNA library of specially or highly expressed genes. The white clones were screened and identified by modified differential screening technique in which the probes were labeled with digoxin/deoxyuridine 5'-triphosphate. Positive clones were sequenced and compared with the known sequences in the public databases of GenBank/EMBL/DDBJ using NCBI BLAST for homology analysis. The unknown fragments were then submitted to GenBank.

RESULTS

A subtractive library of differentially expressed genes for the family with cases of familial acute myelocytic leukemia was constructed successfully. After differential screening, 28 effective sequences were confirmed to be positive differentially expressed gene fragments and were submitted to GenBank for homology analysis. Then 17 sequences were identified as a part of known genes, and the rests were verified as unknown fragments.

CONCLUSION

SSH is an effective approach to isolate differentially expressed genes. Some genes related to cell proliferation and differentiation has been acquired by SSH. Eleven novel expressed sequence tags related to library have been successfully isolated and accepted by GenBank.