Development of Avian Transgenic System

本项目从1992年开始，为建立动物基因表达与调控领域研究人员所要求的禽类转基因体系，通过1992-1994三个财政年度的研究，取得了以下成果：除建立了完整的鸡受精卵体外培养技术外，还建立了类似的鹌鹑受精卵体外培养技术。这项技术可以将Rize用于成人受精卵的体外发育，效率超过25%。2.利用这一完整的禽受精卵体外培养技术，在胚胎期将囊胚层细胞导入其他胚胎，获得了具有嵌合性腺的嵌合鸡。此外，我们还成功地培育出嵌合F1鸡。3.由于蛋黄的大小和光密度，观察禽蛋的卵核是非常困难的。通过电视摄像机和特殊布置的照明系统相结合，建立了检测禽受精卵核区的改进方法，使在核区微量注射待转基因基因成为可能。4.为了提高基因重组到宿主基因组的效率，我们多次尝试将重组的基因与逆转录病毒的整合酶共注射到宿主基因组中，并取得了预期的效果，为建立禽类转基因体系奠定了基础。

This project started from 1992 in order to establish the avian transgenic system which has been required by researchers in the field of gene expression and regulation in animal in situ, ant the following results have been established through studies conducted for three fiscal years from 1992 to 1994.1.In addition to the complete in vitro culture technique for chicken fertilized eggs, the similar technique for quail fertilized eggs has been established. This technique can give rize to in vitro development of fertilized eggs to adults with more than 25% of efficiency. 2.Using this complete in vitro culture technique for avian fertilized eggs, chimeric chickens with chimeric gonads have been produced by introducing blastoderm cells to other embryos at blastoderm stage. In addition, we have succeeded in producing chimeric F1 chickens. 3.It is highly difficult to observe egg nucleus of avian eggs because of their size and optical density owing to yolk. By combining TV-camera and specifically arranged illumination system, improved method to detect nuclear area of avian fertilized eggs has been established, and now micro-injection of genes to be transfected to in the nuclear area has become possible. 4.To improve the efficiency of recombination of transfected genes to the host genom, it has been repeatedly tried to co-inject the gene recombined with retroviral vector together with the integrase derived from retrovirus, and prospective results have been obtained.Thus the fundamental basis required for establishement of avian transgenesis system has been obtained through this project.