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Author:
Kang, H.   Sung, D.  


Journal:
Antiviral Research


Issue Date:
1998


Abstract(summary):

In an attempt to develop efficient way to inhibit the propagation of plant viruses, antisense RNA and ribozyme targeted to the frameshifting site of the plant viruses were designed and their activities were tested using the luciferase reporter system. A 17mer RNA complementary to the RNA sequence including the frameshifting site of the Barley Yellow Dwarf Virus (BYDV) completely arrests the translation of luciferase gene. This inhibitory effect is dependent on the concentration of the antisense RNA. A 36mer hammerhead ribozyme was designed to cleave the RNA at the frameshifting site. Translation of the luciferase gene is arrested by the ribozyme. In order to test the efficacy of the antisense and ribozyme in vivo, a yeast expression system using pYES2 vector was designed. The genes coding the antisense and/or ribozyme were cloned into the Pvu II / Sph I site of the pYES2, which is expressed under the control of the GAL1 promoter. The pYES2 vector was co-transfected into the yeast with the pAS2-1 vector carrying the frameshifting assay cassette of the BYDV. Analysis of the protein products by Western Blotting reveals that antisense and ribozyme completely inhibit the translation of the messenger RNA beyond the target site. Our results indicate that frameshifting site in viruses is a good target site for antisense and ribozyme, which is a potential way to inhibit the viral propagation.


Page:
A91


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