摘要
采用显微注射法将含有鲤鱼 β actin基因启动子的草鱼生长激素基因“全鱼”基因pCAgcGHc转入异源四倍体鲫鲤 ,然后使其自交得到转基因异源四倍体鲫鲤F1,对 15 0日龄F1体重和体长进行检测 ,可明显看见转基因异源四倍体鲫鲤F1的生长优势 ;取F12 0尾 ,提取尾鳍基因组DNA ,采用合适的引物 ,PCR方法检测转基因异源四倍体鲫鲤F1是否含有外源生长激素基因 ,结果 15 0日龄F1阳性率达到 90 % ,且有些雄性个体可以挤出少量精液 ,而普通 15 0日龄异源四倍体鲫鲤无此现象。
The tetraploid fish has been developed by assortative breeding the hybrids of Carassius auratus red var.(♀)×cyprinus carpio(♂),which has the stable genetic characters and can reproduce themselves. An 'all fish'recombinant DNA construct ( pCAgcGHc )containing common carp β actin gene promoter and cDNA for grass carp (Ctenopharyngodon idella)growth hormone gene was introduced into fertilized eggs of the allotetroploid fish through microinjection as soon as artificial insemination was done. Artificial insemination was carried out between the female and the male transgenic allotetraploid fish which contain the 'all fish'recombinant DNA construct ( pCAgcGHc )and are the biggest in the size. Fifty F 1 samples of transgenic allotetraploid fish of 150 days and 50 allotetraploid fish (regarded as the control)were chosen, and the weight and the body length of each were measured, the results showed that F 1 of transgenic allotetraploid fish of 150 days had obvious growth dominance compared with the control. Genomic DNA of tail fin was extracted from 20 F 1 of transgenic allotetraploid fish of 150 days and the control. Proper primers were introduced to check whether the sample had the transgene. Pa, the upstream primer, is located in β actin promoter, and Pg, the downstream primer, is located in growth hormone cDNA for grass carp (gcGHc). The transgene was detected in 90% F 1 of transgenic allotetraploid fish in tail fin DNA by polymerase chain reaction (PCR)amplification. Sperm could be squeezed out from a few F 1 of transgenic allotetraploid fish of 150 days, however, this phenomenon did not exist in the controls. The importance of forming the pure line of transgenic allotetraploid was elucidate in the paper.
