全鱼基因的构建及其在鲫鱼体内的整合与转录

Reconstruction of cMTsGH Gene and Its Integration and Translation in Crucian Carp (Carassius aumtus)

利用PCR技术删除大麻哈鱼生长激素基因的启动序列 ,通过基因重组构建出全鱼基因 (鲤鱼MT启动子 大麻哈鱼生长激素基因 ) ;以融合全鱼基因为外源基因 ,通过显微注射方法将其线性片段导入鲫鱼受精卵内 ,研究其整合与转录效率。结果表明 ,全鱼基因在鲫鱼基因组中的整合率为 36 4 % ( 1 6 4 4) ,对转基因阳性鱼的RNA样本进行Northern印迹杂交检测 ,转录率为 2 5 % ( 1 4 )。因此 ,该全鱼基因可以作为转基因鱼研究和应用的外源基因。

The promoter sequence of growth hormone gene of trout salmon was deleted by PCR,and a reconstructed gene cMTsGH was produced by combing common carp metallothionein promoter and growth hormone gene of trout salmon. The linear DNA fragment of the gene was microinjected into the fertilized eggs of crucian carp, and the rates of integration and translation were examined. The integration rate was 36.4%(16/44) in crucian carp genome. Analysis of RNA samples of the positive transgenic fish by Northern blot showed that the translation rate of the gene was 25%(1/4). It is concluded that cMTsGH, the gene reconstructed by combing common carp metallothionein promoter and growth hormone gene of trout salmon, can act as a foreign gene for the study and production of transgenic fish.