摘要
以含大肠杆菌β-半乳糖苷酶基因,及人工合成启动子的三种转移载体质粒:pSVI^+G,pSVI^-G和pSWVI^-G,重组了三株带人工合成启动子、驱动β-半乳糖苷酶基因的粉纹夜蛾核型多角体病毒TnNPV-SVI^-G,TnNPV-SVI^+G及TnNPV-SWVI^-G。其中TnNPV-SVI^+G形成多角体,TnNPV-SWVI^-G保留了多角体启动子。它们为遗传上稳定、均一的病毒,接种感染草地夜蛾细胞后,25小时可测到β-半乳糖苷酶的活性,50小时达最高,80小时后开始下降。实验表明:保留野生型多角体启动子,对合成启动子表达外源基因有增强作用,但保留启动子与保留多角体基因,则有抑制人工合成启动子表达外源基因的作用。
Recombinant Trichoplusia ni NPV designated TnNPV-SVI^-G,TnNPV-SWVI^-G and TnNPV-SVI^+G respectively were constructed.All these recombinants contain a passenger gene,Eschorichia coli β-galactosidase(β-gal),under the control of a synthetic promoter based on the conserved sequences of Autographa californica NPV late promoters.Additionally,both TnNPV-SWVI^- G and TnNPV-SVI^+G contain the polyhedrin promoter,but minus and plus the coding region respectively in opposite orientation to the synthetic promoter and β-gal gene.Results of β-gal activity determ(?)nations indicated that the expression of β-gal gene is temporally controlled beginning late in infection(25 h.p.i.).A maximal level of expression was achieved by 56 h.p.i.and the β-gal activity declined after 80 h.p.i. Comparison of β-gal expression levels in S.frugiperda cells infected with TnNPV-SVI^-G,TnNPV-SWVI^-G and TnNPV-SVI^+G respectively showed that insertion of the polyhedrin promoter alone had positive effects on the synthetic promoter resulting in increased β-gal expression while the polyhedrin promoter plus the coding sequence had negative effects leading to decreased expression. The advantages of TnNPV-SVI^+G system in foreign gene expression and its possible use in pest control were discussed.
出处
《中山大学学报（自然科学版）》
CAS
CSCD
1990年第3期136-141,共6页
Acta Scientiarum Naturalium Universitatis Sunyatseni
关键词
杆状病毒
合成启动子
基因
表达
baculovirus
synthetic promotor
β-galactosidase gene
