中华蜜蜂Apis cerana cerana是一种真社会性昆虫,也是我国重要的经济昆虫。本实验目的是为了检测精子是否可以作为载体将外源egfp基因介导转入中华蜜蜂。首先将雄蜂精子与线性化的质粒DNA共浴,然后通过人工授精技术将精子导入处女王,再对实验蜂群后代进行分析。结果显示EGFP蛋白在一群实验组蜂的1~2日龄小幼虫中表达较强,能检测到0.01%~0.02%荧光阳性小幼虫个体;通过PCR和RT—PCR技术分析,证实转入的外源egfp基因获得表达。实验结果表明精子载体法能够用于中华蜜蜂外源基因的转移和表达。
The Chinese honeybee, Apis cerana cerana is a eusocial insect and one of the most important economical insects in China. The objective of this study was to transfer the exogenous egfp gene through spermatozoa in A. cerana cerana. The drone sperms were mixed with foreign linearized DNA, which were then transferred into the oviducts of a virgin honeybee queen with the technique of artificial instrumental insemination. The descendants of the experimental colonies were analyzed. Green fluorescence was observed in 1- to 2-day-old larvae from a colony of the experimental group, and the positive rate was 0.01% - 0.02%. PCR amplification of genomic DNA and reverse transcription-polymerase chain reaction confirmed the transfer and expression of egfp gene. The results indicate that sperm-mediated transformation can be applicable to transfer and express exogenous genes in A. cerana cerana.