Iridovirids　are　a　group　icosahedral　viruses　containing　linear　double-stranded　DNA,　and　mainly　infect　invertebrates　and　poikilothermic　vertebrates.　Cherax　quadricarinatus　iridovirus　(CQIV)　is　a　new　species　of　the　family　Iridoviridae　and　can　cause　high　mortality　in　shrimps.　In　CQIV　genome,　there　are　25　conserved　genes　and　the　putative　products　are　involved　in　several　viral　processes.　In　this　study,　three　core　protein　including　CQIV032R,　CQIV-125R　and　CQIV-160L　were　identified　to　interact　with　CQIV-038R　by　yeast　two-hybrid　(Y2H),　and　the　interaction　between　CQIV-038R　and　CQIV-125R　was　further　confirmed　by　co-immunoprecipitation　(Co-IP)　assays.　In　the　expression　system,　EGFP-038R　and　mCherry-125R　were　colocalized　in　the　cytoplasm　when　co　expressed　in　Sf9　cells.　Moreover,　silencing　the　expression　of　038R,　125R　or　both　of　these　two　proteins　respectively　in　C.　quadricarinatus　cells　by　small　interfering　RNAs　showed　significantly　inhibit　CQIV　replication.　Collectively,　we　identified　the　interaction　between　038R　and　125R,　and　demonstrated　they　are　essential　for　CQIV　replication.