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In recent years,much effort has been made in identifying microRNA (miRNA) genes from mammals,insects,worms,plants,and viruses.Continuing the search for more miRNA genes is still important but difficult.This paper presents a computational strategy based on comparative genomics analysis.The algorithm was used to scan four invertebrate genomes,Drosophila melangoster,Bombyx mori,Apis mellifera,and Anopheles gambiae,which are either model organisms or medically/economically important insects.99 new miRNA genes were predicted from the four insect species which can be grouped into 17 miRNA gene fami-lies,of which 10 of the miRNA families are insect-specific.Sequence similarity analysis showed that 16 of the newly predicted insect miRNAs belong to the K-box,GY-box,and Brd-box miRNA families which are impor- tant participators in Notch-related pathways,To test the validity of the algorithm,39 predicted insect miRNA genes from D.Melangoster and A.mellifera were selected for further biological validation.34 (87%) pre- dicted miRNA genes' transcripts were successfully detected by reverse transcription-polymerase chain reac- tion experiments.Thus,this strategy can be used to efficiently screen for miRNA genes conserved cross species.