中文摘要：

鳞翅目昆虫粉纹夜蛾（Trichoplusiani）的PiggyBac（PB）转座子已用于模式生物小鼠的转基因及插入诱变研究，目前，该转座子在养殖鱼类中的转基因效率如何还不清楚。构建了带PiggyBac转座子左臂、右臂、EF1a启动子和绿色荧光蛋白（eGFP）编码框的pPBs—EF1a-eGFP供体质粒。以50ng／乩供体质粒和100ng／μL体外转录的PB转座酶mRNA共同显微注射入团头鲂（Megalobrama amblycephala）1～2细胞期受精卵中，团头鲂eGFP的荧光表达率可达58．26％，PCR检测结果显示，该转座系统在团头鲂成鱼基因组中的整合效率为53．04％。表明PiggyBac转座子可高效介导基因在团头鲂基因组中的插入，为进一步开展团头鲂插入诱变研究奠定了基础。

英文摘要：

PiggyBac （PB） transposon is derived from Trichoplusia ni of lepidopteran and has been widely used in transgenic and insertion mutagenesis studies in mouse. Currently, relevant research about PiggyBac transposons used in farmed fish has not yet been reported. To study insertion efficiency of PiggyBac transposon in the genome of M. amblycephala, we built pPBs-EFI~-eGFP donor plasmid with PiggyBac transposon left and right arms, EFlc~ promoter and eGFP gene, then co-injected with PiggyBac transposase mRNA into the 1 -2 cell stage fertilized eggs of M. amblycephala. The concentrations of donor plasmid and transposase mRNA were 50 ng/μL and 100 ng/μL respectively. The eGFP fluorescence expression rate was 58.26%. In adult fish, PCR results demonstrated that integration efficiency of PiggyBac transposition system was 53.04% in M. amblycephala genome. Our data suggest that PiggyBac transposon can efficiently mediate gene insertion in M. amblycephala, which could be used in insertional mutagenesis in M. amblycephala.