Establishment of chicken transgenic technology using embryonic stem cells.

本课题主要研究利用胚胎干细胞建立鸡转基因技术。我在基因克隆、重组蛋白和抗体的构建、胚胎干细胞的细胞培养、靶向载体的构建等方面进行了实验，得到如下结果：(1)克隆了鸡的哺乳动物白血病抑制因子（LIF）和白细胞介素(IL)-6同源基因cdna及鸡的LIF和IL-6的表达。此外，我还开发了针对这些因素的抗体。我的结论是，鸡的LIF能够维持鸡胚胎干细胞培养处于未分化状态。(2)从抗鸡LIF抗体实验来看，鸡LIF激活STAT3对维持鸡ES细胞的未分化状态有显著作用。(3)确定了对鸡胚胎干细胞进行转基因和筛选（阳性和阴性）的条件。(4)克隆了鸡δ-晶体蛋白和卵黄蛋白原基因的基因组dna，构建了两个靶向载体。我的发现应该有助于这项技术的发展。

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.