外源生长激素基因在蓝太阳鱼中的整合、表达和遗传

Integration, expression, and inheritance of foreign growth hormone gene in Lepomis cyanellus

通过基因重组, 将石斑鱼生长激素基因编码序列克隆到鲤鱼βactin基因启动子下游, 构建了“全鱼”生长激素基因表达载体pCAecGHc。采用显微注射法, 研制出转“全鱼”生长激素基因蓝太阳鱼。经过PCR、PCR Southern杂交、RT PCR等技术对转植基因在转基因蓝太阳鱼中的整合和表达情况进行了检测。结果表明:转植基因在两批次P0 转基因蓝太阳鱼中的整合率为5 .60%和12. 26%, 并在转基因鱼中得到了正确表达, 表现为嵌合性表达。对转基因蓝太阳实验鱼进行了对照养殖实验, 初步显示转基因蓝太阳鱼具有较快的生长表型效应, 比对照组生长速度快20%-40%左右。应用近交策略培育出了两个转基因蓝太阳鱼F1 品系, 检测表明, 转植基因在两个品系的整合率分别为22 .03%和40 .8%。结果表明: 转植基因通过性腺传递给了子代, 同时也证实转基因P0 代的生殖腺为转植基因的嵌合体。

The coding sequence of the growth hormone gene of the grouper Epinephelus coioides was cloned under the regulation of carp β-actin gene promoter and enhancer, so an “all-fish” recombinant construct pCAecGHc was constructed. Then the construct was microinjected into eggs of Lepomis cyanellus and the transgenic Lepomis cyanellus was produced. Through the detection methods of PCR, PCR-Southern blot, and RT-PCR, the integration and expression of transgene in host fish were determined. The results showed that the integration ratio of transgene was 5.60%-12.26%, and that the transgene was correctly transcribed and expressed in mosaic fashion in host fish. Preliminary culture trials showed that P 0 of transgenic Lepomis cyanellus grew faster than the non-transgenic siblings with 20%-40%. Two F 1 lines of transgenic Lepomis cyanellus were obtained by inbreeding and the integration profile of transgene in F 1 was investigated. The positive rate of transgene in two F 1 lines was 22.03% and 40.8% respectively. The results suggested that the gonads of P 0 transgenic Lepomis cyanellus were transgenic mosaics and also provided convincing evidence that the transgene CAecGHc was transmitted to the progeny through the germline.