Promoter studies of a polydnavirus gene from Cotesia rubecula (Hym: Braconidae).
The Cotesia rubecula polydnavirus gene, CrV1, is expressed in a highly transient fashion. Within four hours after egg deposition and virus infection, tissues of the host caterpillar, Pieris rapae, express high levels of the transcript. Twelve hours after infection no transcripts are visible. We have previously shown that the CrV1 secreted protein is mainly produced in host haemocytes. In haemocytes, immune functions such as phagocytosis and cell spreading are abolished by destabilization of the cell cytoskeleton. To test whether the observed down-regulation of CrV1 transcripts is mediated by transcriptional control or by other factors, such as the disruption of cytoskeleton in CrV1-inactivated cells, we cloned the promoter and the 3' untranslated region of the CrV1 gene to study CrV1 expression. The promoter region of the CrV1 gene was cloned into baculovirus expression systems along with the CAT reporter gene. Molecular analyses showed that the CAT gene under the control of CrV1 promoter is expressed as early as 2 h post infection and continues until late phase of infection suggesting that down-regulation of CrV1 expression in host haemocytes is perhaps mediated by post-transcriptional mechanisms.
Asgari
S
S
Department of Applied and Molecular Ecology, Adelaide University, Glen Osmond, Australia. sassan.asgari@adelaide.edu.au
Schmidt
O
O
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry / Q000302:isolation & purification; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D018071:Polydnaviridae / Q000235:genetics*; D011401:Promoter Regions, Genetic; D018411:Spodoptera; D014863:Wasps / Q000821:virology*
D004279:DNA, Viral; D015500:Chloramphenicol O-Acetyltransferase