基于piggyBac转座子转hGM-CSF基因家蚕的研究

Research on hGM-CSF Transgentic Silkworm With piggyBac Transposon

将由家蚕核型多角体病毒IE-1基因启动子控制下的hGM-CSF基因克隆到p igA3GFP载体中,构建了家蚕转基因载体p igA3GFP[IE-GMCSF],利用压力渗透法和精子介导法将其与辅助质粒helper p igA3一起导入家蚕蚕卵,获得产生绿色荧光的家蚕,次代产生荧光蚕的比例分别为0.17%,0.15%。将次代荧光蚕与正常蚕交配后代(G1)的荧光蚕个体再相互杂交,连续进行多代选育,获得了稳定遗传的转hGM-CSF基因家蚕品系。

To approach hGM-CSF transgentic silkworm Bombyx mori, the victor pigA3GFP [IE-GMCSF] was constructed by cloning hGM-CSF with IE-1 promotor of BmNPV into pigA3GFP vector, and transported into eggs by pressure penetrating and sperm mediating together with the nonautonomous helper pigA3 which encodes piggyBac transposase. The ratio of green fluorescent larvae is 0.17% and 0.15% respectively in G0 generation. G1 was generated by mating G0 green fluorescent silkworms with normal silkworms, then hybridization was carried out among the individuals of G1 green fluorescent larvae. After continuously several hybridizations among green fluorescent silkworms, the steady transgenetic hGM-CSF silkworm strain was obtained.