电穿孔作用将外源基因导入中华蜜蜂卵的研究

Introduction of foreign DNA into Apis cerana cerana eggs by electroporation

将中华蜜蜂卵与带有绿色荧光蛋白基因的质粒DNA混合,经电穿孔作用后,人工孵化蜜蜂卵,并对孵化的幼虫进行检测。结果筛选出一组较理想的中华蜜蜂卵电穿孔参数(750V.cm-1,100μs,1);通过荧光检测,电击卵孵化的幼虫体内可以表达EGFP基因,阳性表达率最高可达1.05%,且外源DNA转染早期卵的效率显著高于晚期卵。结果表明电穿孔作用成功地将外源基因引入中华蜜蜂卵内,质粒DNA转染中华蜜蜂卵的效率与卵的生长时期有关。

The Apis cerana cerana eggs were electroporated after they were mixed with plasmid DNA encoding enhanced green fluorescent protein （EGFP） gene. The electroporated eggs were artificially fertilized, and the hatched larvae were analyzed. Here we found a group of optimizing parameter for the Apis cerana cerana eggs electroporated （750 V·cm^-1,100μs,1 ）. The express of EGFP gene was proved in the hatched larvae with the fluorescence microscope. The highest percentage of hatched larvae with foreign genes was 1.05 %. The efficiency was more significantly increased for foreign DNA transfection in early eggs than late eggs. The results showed that the foreign gene had been successfully introduced into the Apis cerana cerana eggs by electroporation, and the efficiency of plasmid DNA transfection in Apis cerana cerana eggs was related to the grow period of eggs.