Combining Suppression Subtractive Hybridization(SSH)and cDNA Microarray for Rapid Identification of Differentially Expressed Genes From Influenza Virus Infected Host Cells
Current antiviral drugs mainly target to viral proteins,such as reverse transcriptase and proteinase.However,virus protein-targeted drugs usually bring alout resistance.On the other hand,the number of targets for virus is limited due to the small coding capacity of viral genome.So,an alternative strategy for antiviral drug target research is to screen and validate the host-derived genes and/or proteins that are essential for the virus to complete its replication.To find the potential cellular antiviral targets which are the differentially expressed genes during influenza virus infection,subtractive cDNA library was constructed by suppression subtractive hybridization(SSH)with influenza virus A/Lufang/93-9(H3N2)infected-MCDK cells as tester and the mock-infected MDCK cells as driver.Then cDNA microarray containing 768 cDNA fragments and control genes was used to further identify the differentially expressed genes in cDNA library.It showed that the length of inserts in subtractive cDNA library were between 250～1000bp.18 differentially expressed clones were selected by cDNA microarray to be novel ESTs as no functional clues were associated with them by bioinformatic analysis.These novel ESTs differentially expressed in the course of influenza virus infection would provide a good foundation for further finding of host-derived antiviral target and/or diagnostic biomarker,and also using for dissection of the molecular pathogenesis of influenza virus infection.