显微介导河蟹总DNA导入镜鲤的检测与分析

摘要

In this study,total DNA of Chinese mitten handed crab (Eriocheir sinensis) was injected into mirror carp (Cyprinus carpio) by micro-injected distant hybridization technique,and the exogenous DNA in mirror carp micro-injected Chinese mitten handed crab was detected by AFLP molecular marker technology.In the 30 pairs of AFLP primers,3 pairs of primers have amplified the gene fragments of donor,indicating that there is the micro-injected Chinese mitten crab gen found in only crab.The results showed that,cloned and sequenced amplification products revealed that there were transgene fragments in all the positive three scales of 6 positive micro-injected mirror carp.The findings verified that the total DNA of crab was injected into three scales genome at molecular and gene levels,providing a new strategy for the application of micro-injected exogenous total DNA.%利用显微介导远缘杂交技术将河蟹总DNA直接导人镜鲤受精卵内,再利用AFLP分子标记技术检测显微介导河蟹基因镜鲤的外源DNA.在30对AFLP引物中,有3对引物扩增出供体基因片段,即在显微介导河蟹基因镜鲤中均有和河蟹基因相同而对照镜鲤没有的条带.对6尾阳性显微介导河蟹基因镜鲤的扩增产物进行回收、克隆和测序验证.结果表明:阳性显微介导河蟹基因在镜鲤中均含有供体基因目的片段.本研究在分子和基因水平上验证了河蟹总DNA可通过显微介导方式整合到镜鲤基因组中,为显微介导外源总DNA转化技术的应用提供新的思路.