Establishment of chicken transgenic technology using embryonic stem cells.

在这个项目中，我研究了利用胚胎干细胞建立鸡的转基因技术。通过基因克隆、重组蛋白和抗体的研制、胚胎干细胞的细胞培养和靶向载体的构建等实验，获得了以下结果：(1)克隆了鸡白血病抑制因子(LIF)和白介素6(IL-6)的同源基因，并在鸡LIF和IL-6中进行了表达。此外，我还开发了针对这些因素的抗体。从抗鸡LIF抗体的实验来看，鸡LIF激活STAT3对维持鸡ES细胞的未分化状态具有重要意义。(3)确定了鸡ES细胞转基因和筛选(阳性和阴性)的条件。(4)克隆了鸡δ-晶体蛋白和卵黄蛋白基因的基因组DNA，并构建了两个靶向载体，为该技术的发展奠定了基础。

In this project, I investigate to establish chicken transgenic technology using embryonic stem cells. I obtained the results below experimenting on the gene cloning, development of recombinant proteins and antibodies, cell culture of embryonic stem cells and construction of targeting vectors.(1)I cloned the cDNAs of the chicken homologue of mammalian leukemia inhibitory factor (LIF) and interleukin (IL)-6 and expression of chicken LIF and IL-6. In addition, I developed the antibodies against these factors. I conclude that chicken LIF is able to maintain chicken ES cell cultures in the undifferentiated state.(2)From the experiment of anti-chicken LIF antibody, activation of STAT3 by chicken LIF was significant for maintaining the undifferentiated state of chicken ES cells.(3)The conditions of transgene and selection (positive and negative) against chicken ES cells were decided.(4)I cloned the genomic DNAs of chicken δ-crystallin and vitellogenin genes, and constructed two targeting vectors.My findings should contribute to the development of this technology.