Screening and cloning of differential expressed genes in child acute lymphocytic leukemia by suppression subtractive hybridization

Objective To clone and identify genes differentially expressed in child acute lymphocytic leukemia ( ALL ) by suppression subtractive hybridization. Methods A subtracted cDNA library of ALL was constructed by suppression subtracted hybridization ( SSH) method. After screening,the clones that subtracted library were selected to identify by cloney PCR and DNA sequencing. Nucleic acid homologies were performed using the BLAST program. Results Random analysis of 60 clones from subtracted cDNA library showed about 200-600 bp inserts confirmed by colony PCR,and 20 clones were confirmed by sequence analysis and blast analysis. The 18 clones encoding 17 genes were highly homologous with the partial sequences of known genes,the coincidence rates were 98% -100% . These genes were related with cell cycles,metabolism,and tumor development closely. Two new genes were found. Conclusion It was concluded that a subtractive cDNA library related to pediatric ALL was constructed successfully,which laid foundation for screening related genes in children ALL.