【摘要】 生态安全性是转基因鱼走向市场的瓶颈,通过转基因四倍体鱼同转基因二倍体鱼杂交获得不育的转基因三倍体鱼是解决该问题的有效途径之一.本研究构建了青鱼β-actin基因启动子和青鱼生长激素(GH)基因精确连接的"全鱼"基因pbcAbcGHc;并采用显微注射法将pbcAbcGHc导入异源四倍体鲫鲤受精卵.对照养殖结果表明,150日龄的转基因异源四倍体鲫鲤原代(P0)的体重及体长明显大于对照组.选择60尾P0代转基因异源四倍体鲫鲤,采用PCR方法检测出外源青鱼GH基因在P0代转基因四倍体尾鳍基因组DNA中的整合率为90%;对20尾雄性P0代转基因四倍体精液样本的PCR检测发现,13个样本具有外源青鱼GH基因的整合.在一尾生长速度显著的P0代转基因四倍体鲫鲤的肌肉、肝脏、肾脏和卵巢组织中可检测到外源青鱼GH基因的转录.本研究成功获得了具有明显生长优势的P0代转青鱼GH基因异源四倍体鲫鲤,为建立转青鱼GH基因异源四倍体鲫鲤纯系和研制不育的转基因三倍体鱼奠定了基础.更多还原

【Abstract】 The ecological risk is the "bottleneck" for commercialization of transgenic fish.The sterile transgenic triploid fish derived from hydrating the transgenic tetraploid fish and transgenic diploid fish is one of the keys to solve this problem.The "all fish" gene construct(pbcAbcGHc) containing Black Carp B-actin gene promoter and cDNA of growth hormone(GH) gene from the same species was introduced into the fertilized eggs of the allotetroploid fish through microinjection.The contrast cultivation results show that the weight and body length of P0 of the transgenic allotetraploid fish of 150 days were much larger than those of the controls.PCR assay was applied among 60 transgenic allotetraploid fish of 150 days and the integration ratio of the exogenous transplant gene in the genome of the caudal fin is 90%.The transplant gene could be detected in 13 of 20 samples from the sperm of the male transgenic tetraploid fish.The RT-PCR result demonstrates that the transcription of exogenous GH gene could be found in the muscle,liver,kidney and oval of the biggest transgenic allotetraploid fish.This study has developed P0 of the Black Carp GH gene transgenic tetraploid fish with outstanding growth rate,which made the solid foundation for the pure line establishment of the transgenic tetraploid fish and for developing the sterile transgenic triploid fish.更多还原