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采用显微注射法将人α干扰素(HuIFNα)重组基因转入草鱼Ⅰ～Ⅱ细胞期的受精卵,然后对转化培育出的草鱼个体提取血液总DNA进行PCR和Southern杂交检测及ELISA检测,以研究重组基因在受体基因组上的整合和表达情况。实验结果显示:HuIFNα重组基因能整合在草鱼基因组上,但其整合是随机的,整合位点没有固定区域,而且整合在受体草鱼基因组上的HuIFNα基因的表达率较低,表达水平在个体间有很大差异。大部分转基因个体外源基因表达水平集中在39～74pg/mL之间,表达最高个体HuIFNα浓度为1450pg/mL。
The recombinant human interferon alpha (HuIFNα) gene was transferred into the fertilized eggs of stage Ⅰ ~ Ⅱ of grass carp by microinjection method. Then the total DNA of blood was extracted from the transformed grass carp. PCR and Southern blot were carried out to detect To study the integration and expression of recombinant genes on the receptor genome. The results showed that the recombinant HuIFNα gene could be integrated into the genome of grass carp, but its integration was random. There was no fixed region in the integration site. The expression of HuIFNα gene was lower in the grass carp genome, There are big differences. Most of the genetically modified individuals express the level of exogenous gene concentrated between 39 ~ 74pg / mL, the highest expression of individual HuIFNα concentration of 1450pg / mL.