中文摘要：

为探索草鱼出血病防治新技术,实验采用衣壳蛋白靶向灭活（capsid-targeted viral inactivation,CTVI）策略,利用Tgf2转座子元件,构建了带爪蟾EF1α或鲤热休克蛋白70两种不同启动子的CTVI转基因质粒p Tgf2-EF1α-VP3-SN或p Tgf2-Hsp70-VP3-SN。该2种质粒均包含GCRV衣壳蛋白VP3与金黄色葡萄球菌核酸酶（Staphylococcus aureus nuclease,SN）融合表达阅读框。通过将2种CTVI转基因质粒与体外合成的Tgf2转座酶5＇加帽mRNA共同显微注射入草鱼1~2细胞期受精卵,获得了带2种不同启动子的CTVI转基因草鱼群体。PCR和测序结果显示,2种转基因阳性草鱼基因组中均含有外源GCRV衣壳蛋白VP3与SN基因片段,阳性率分别为40.2%和37.0%,表明Tgf2转座子已成功介导CTVI融合表达基因整合到草鱼基因组中。本实验共获得了120尾P0CTVI转基因个体,为将来构建抗出血病草鱼新品系奠定了基础材料。

英文摘要：

To explore new methods to prevent the grass carp （Ctenopharyngodon idella）hemorrhage, the capsid-targeted viral inactivation （CTVI） strategy was developed in the present study. By combination with Cyprinus carpio heat shock protein 70 or Xenopus EF1 a promoters, two different transgenic plasmids pTgf2- EF1a-VP3-SN and pTgf2-Hsp70-VP3-SN were constructed harboring with efficient Tgf2 transposon element. Both transgenic plasmids contain open reading frame（ORF） fusion with GCRV capsid protein VP3 and Staphylococcus aureus nuclease （SN）. The transgenic grass carp was produced by co-injection pTgf2- EF1α-VP3-SN or pTgf2-Hsp70P-VP3-SN plasmids into 1 -2 cell fertilized eggs with in vitro synthesized Tgf2 transposase mRNA. The PCR and sequencing analysis have proved that the anti-GCRV transgenic systems have successfully been integrated into the genome of grass carp. The transgene positive rates of pTgf2-EF1 et-VP3-SN or pTgf2-Hsp70P-VP3-SN plasmids are 40.2% and 37.0% , respectively. Our results demonstrated Tgf2 transposon can efficiently mediate transgenesis in grass carp for fusion ORF of GCRV capsid protein VP3 and SN. Total 120 transgenic grass carp individuals have been obtained in the present study. The construction of anti-GCRV transgenic P0 will provide the materials for future transgenic breeding to prevent grass carp hemorrhage.