将pGFP-2质粒线性化,利用精子介导,通过人工授精技术导入意大利蜜蜂处女王,然后将授精王介绍到无王群繁殖后代。结果:试验群产生了一群绿色荧光阳性蜜蜂;对阳性蜂群后代分析发现,1-2日龄幼虫能检测到荧光,提取蜜蜂DNA进行PCR扩增得到预想的片断,通过RT-PC分析,从转录水平上证实转导的外源基因获得表达。结论:以绿色荧光蛋白(GFP)基因为报告基因,通过精子介导法能够生产克隆转基因蜜蜂。
The plasmid DNA was linearized and introduced with sperm during the instrumental insemination of virgin queen, then these inseminated queens were introduced into queenless colonies. Results : a green fluorescence - positive colony was produced, where green fluorescence was observed for 1 to 2 - day - old lar- vae, the predicted fragment was isolated by means of PCR amplification of genomic DNA and the expression of transferred genes was confirmed at the transcriptional level by reverse transcription - polymerase chain reaction. Conclusion: using green fluorescent protein (GFP) as a reporter gene, transgenic honey bees could be established by SMGT.