Anticarsia gemmatalis MNPV IE-1 trans regulatory protein, studies on its activity

BILEN MARCOS; MARIA JULIA OUSSET; GOÑI SANDRA; BELAICH MARIANO; LOZANO MARIO ENRIQUE; GHIRINGHELLI DANIEL

Villa Carlos Paz

Congreso; XXXVII Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2001

AgMNPV is today the most widely used biopesticide in more than one million hectares of soybean in Brazil. It kills the velvetbean caterpillar Anticarsia gemmatalis (plague in Brazil and Argentina), but depending on the age and susceptibility of the host, this virus may take up to two weeks to kill the insect. Taking into account these facts, our aim is to study the general characteristics and regulatory mechanisms involving immediate early promoters in the viral cycle. The ie-1 gene codes for a regulatory protein that has been involved in transcriptional activation of other genes, in its own transcriptional regulation and to participate in the viral DNA synthesis. In this work, we shows studies on the role of IE-1 protein in the transcriptional regulation of its own gene, in transient experiments using truncated versions of the ie-1 promoter region driving the expression of a reporter protein (E. coli lac Z). A plasmid carrying the complete locus of ie-1 were cotransfected with the different constructs in UFL-AG-286 insect cells, to assess the time course and levels of Lac Z expression using a ƒÒ-galactosidase activity assay. On the other hand, the ie-1 gene was expressed in the pET-22b(+)/E. coli BL21-DE3 prokaryotic system, in order to generate policlonal antibodies that could be used in future experiments