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KMID : 0578319970070030326
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Molecules and Cells
1997 Volume.7 No. 3 p.326 ~ p.334
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Ribozyme Mediated Targeting of Cucumber Mosaic Virus RNA 1 and 2 in Transgenic Tobacco Plants
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Kwon, Chang Seob
Chung, won Il/Paek, Kyung-Hee
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Abstract
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Hammerhead ribozymes have been extensively used to inhibit the expression of cellular genes or viral genes mainly in the animal study. In this study, we designed a ribozyme targeting the conserved leader sequences of cucumber mosaic virus (CMV)RNA 1 and 2. The ribozyme, with asymmetric lengths of flanking complementary regions, cleaved a model substrate RNA efficiently at 26℃ as well as at 37℃ or 50℃ in vitro. And the ribozyme encoding sequence was introduced into tobacco plants and expressed with the CaMV 35S promoter and 3´ NOS terminator in a monomeric type (pBIR1), tandemly repeated type (pBIR3), and cotranscriptionally combined type (pRokR) with 2.2 copies of I_(17)N satellite RNA. Virus challenging experiments in F_(1) plants of respective transformants with CMV-Y showed specific reductions of viral RNA 1 and 2 in comparison with RNA 3 or 4. Although young plants of a three-leaf-stage showed rather similar mild symptom attenuations in all constructions compared to CMV-Y inoculated wild type, fully grown plants showed a differential degree of resistance upon systemic infections of CMV-Y in pRokR, pBIR3 and pBIR1 transformed plants in a decreasing order.
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