Functional and structural characterisation of AgMNPV ie1.
We have located and cloned the Anticarsia gemmatalis multicapsid nucleopolyhedrovirus isolate 2D (AgMNPV-2D) genomic DNA fragment containing the immediate early 1 ORF and its flanking regions. Computer assisted analysis of the complete ie1 locus nucleotide sequence information was used to locate regulatory signals in the upstream region and conserved nucleotide and amino acid sequences. Comparative studies led to the identification of several characteristic protein motifs and to the conclusion that AgMNPV-2D is more closely related to Choristoneura fumiferana defective NPV than to other Group I nucleopolyhedrovirus. We have also shown that the AgMNPV IE1 protein was able to transactivate an early Autographa californica MNPV promoter and its own promoter in transient expression assays. In order to investigate the biological functionality of the ie1 promoter, the ie1 upstream activating region (UAR) was molecularly dissected and cloned upstream of the E. coli lacZ ORF. The results obtained, after transfection of UFL-AG-286 insect cells, leading us to find that the -492 and -357 versions contains sequence motifs important for the level of the lacZ reporter gene expression.
Laboratorio de Ingeniería Genética y Biología Celular y Molecular, Departamento de Ciencia y Tecnología, Centro de Estudios e Investigaciones, Universidad Nacional de Quilmes, Roque Saenz Peña 352, 1876 Bernal, Buenos Aires, Argentina.
Marcos Fabián
MF
Bilen
Marcela Gabriela
MG
Pilloff
Mariano Nicolás
MN
Belaich
Vanina Gabriela
VG
Da Ros
Julio Carlyle
JC
Rodrigues
Bergmann Morais
BM
Ribeiro
Víctor
V
Romanowski
Mario Enrique
ME
Lozano
Pablo Daniel
PD
Ghiringhelli
D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D019677:Artificial Gene Fusion; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D017930:Genes, Reporter; D017874:Immediate-Early Proteins / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D009036:Moths / Q000821:virology; D017924:Nucleopolyhedroviruses / Q000235:genetics* / Q000502:physiology; D012045:Regulatory Sequences, Nucleic Acid; D017422:Sequence Analysis, DNA; D015533:Transcriptional Activation / Q000502:physiology; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D004279:DNA, Viral; D017874:Immediate-Early Proteins; D001616:beta-Galactosidase