Baculovirus-mediated expression of sporamin from sweet potato for insect control
| Autor: | Cheng-Kuo Lin, 林政國 |
|---|---|
| Rok vydání: | 2005 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 93 Genetically modified Autographa californica multiple nucleopolyhedro- virus (AcMNPV), was constructed with insecticidal sporamin gene derived from the storage protein of the sweet potato. Sporamin was expressed by two distinct and temporally regulated promoters. These two promoters were the pag90 early promoter and the very late p10 promoter, resulting in recombinant viruses named vAv/p10-sp and vAc/pag90-sp, respectively. Western blot analysis revealed a detectable amount of sporamin was expressed under the control of the very late promoter, whereas sporamin was weakly detected when the expression was driven by the early pag90 promoter. To evaluate the enhancement of insecticidal activity, larvae were treated with 1.0E+01∼1.0E+08 PIBs/ml wild-type AcMNPV or recombinant virus. The LT50 (median lethal time, LT) values of 2nd∼4th-instar Trichoplusia ni larvae and third-instar Spodoptera exigua larvae were infected with recombinant viruses shorten by 24 hrs as compared to AcMNPV, the cumulative mortality and LC50 (median lethal concentration LC) also revealed that the recombinant viruses were more active than wild-type-virus. Furthermore, the weights of the recombinant-virus-fed larvae were reduced significantly as compared to wild-type-virus fed larvae. Results also showed the indicating a dose-dependent effect. Finally, we have demonstrated that baculovirus-mediated expression of a trypsin inhibitor, sporamin, using two distinct temporally regulated viral promoter improved the insecticidal efficacy. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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