突变型绿色荧光蛋白基因在昆虫细胞和幼虫中的表达

Expression of the Mutant Green Fluorescent Protein Gene in Insect Cells and Larvaae

用细菌—杆状病毒穿梭系统（Ｂａｃ－ｔｏ－Ｂａｃ）将一套含有绿色荧光蛋白基因（ｇｆｐＳ６５Ｔ）和完整多角体蛋白基因的表达盒转座到ＡｃＮＰＶ－Ｂａｃｍｉｄ的Ｔｎ７转座接受位点，从而构建出一种带有ｇｆｐ基因和完整多角体基因的重组杆状病毒，用这一重组病毒感染细胞能稳定地形成多角体，并在紫外光照射下产生强烈的绿色荧光，以重组病毒感染粉纹夜蛾（Ｔｒｉｃｈｏｐｌｕｓｉａｎｉ）幼虫后，在阳光下即可看到发射绿色荧光的幼虫，在手提式长波紫外光照射下绿色荧光更为强烈。

A expression cassette containing the mutant green fluorescent protein gene (gfpS65T) and the intact polyhedrin gene was transposed to the attachment site for Tn7 in AcNPV Bacmid by using the recombinant transposon vector pFGP, thus resulting in a recombinant baculovirus which carries the gfp gene and another copy of polyhedrin gene. The Sf9 cells infected by the recombinant baculovirus can stably produce polyhedra and show strong green fluorescence under ultraviolet light since the gfpS65T gene expression. Concomtantly, Trichoplusia ni larvae inoculated with the recombinant baculovirus show also green under sunlight and appear stronger green fluorescence under irradiation of a hand hold long wavelength ultraviolet light. It is possible that the recombinant baculovirus bearing the reporter gene gfp are able to be useful for studying entomological ecology and baculovirus epidemiology et al .