Interaction of Heliothis armigera Nuclear Polyhedrosis Viral Capsid Protein with its Host Actin
| Autor: | Song-Ya Lu, Yi-Peng Qi, Guo-Qiong Ge |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
DNA
Complementary Insecta Time Factors Green Fluorescent Proteins Molecular Sequence Data macromolecular substances Transfection Polymerase Chain Reaction Biochemistry Green fluorescent protein Viral Matrix Proteins Capsid Two-Hybrid System Techniques Fluorescence microscope Animals Amino Acid Sequence Promoter Regions Genetic Molecular Biology Actin Gene Library Base Sequence biology cDNA library fungi Nuclear Polyhedrosis Virus General Medicine biology.organism_classification Fusion protein Molecular biology Actins Nucleopolyhedroviruses Lepidoptera Luminescent Proteins Microscopy Fluorescence Cytoplasm Capsid Proteins Plasmids Protein Binding |
| Zdroj: | BMB Reports. 35:562-567 |
| ISSN: | 1976-6696 |
| DOI: | 10.5483/bmbrep.2002.35.6.562 |
| Popis: | In order to find the cellular interaction factors of the Heliothis armigera nuclear polyhedrosis virus capsid protein VP39, a Heliothis armigera cell cDNA library was constructed. Then VP39 was used as bait. The host actin gene was isolated from the cDNA library with the yeast two-hybrid system. This demonstrated that VP39 could interact with its host actin in yeast. In order to corroborate this interaction in vivo, the vp39 gene was fused with the green fluorescent protein gene in plasmid pEGFP39. The fusion protein was expressed in the Hz-AM1 cells under the control of the Autographa californica multiple nucleopolyhedrovirus immediate early gene promoter. The host actin was labeled specifically by the red fluorescence substance, tetramethy rhodamine isothicyanete-phalloidin. Observation under a fluorescence microscopy showed that VP39, which was indicated by green fluorescence, began to appear in the cells 6 h after being transfected with pEGFP39. Red actin cables were also formed in the cytoplasm at the same time. Actin was aggregated in the nucleus 9 h after the transfection. The green and red fluorescence always appeared in the same location of the cells, which demonstrated that VP39 could combine with the host actin. Such a combination would result in the actin skeleton rearrangement. |
| Databáze: | OpenAIRE |
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