Establishment of chicken transgenic technology using embryonic stem cells.

利用胚胎干细胞建立鸡转基因技术。

• 批准号: 16580279
• 负责人: HORIUCHI Hiroyuki
• 金额: $ 2.37万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580279/
• 关键词: chicken; LIF; embryonic stem cells

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.

本课题研究利用鸡胚胎干细胞建立鸡的转基因技术。本人在基因克隆、重组蛋白和抗体的研制、胚胎干细胞的细胞培养以及靶向载体的构建等方面进行了实验，取得了以下结果。(1)I克隆了鸡白血病抑制因子（LIF）和白细胞介素（IL）-6同源基因的cDNA，并进行了鸡LIF和IL-6的表达。此外，我开发了针对这些因素的抗体。我的结论是，鸡LIF是能够维持鸡ES细胞培养在未分化状态。（2）抗鸡LIF抗体实验表明，鸡LIF激活STAT 3对维持鸡ES细胞未分化状态具有重要意义。（3）确定了鸡ES细胞的转基因和筛选（阳性和阴性）条件。(4)I克隆了鸡δ-晶状体蛋白和卵黄蛋白原基因的基因组DNA，构建了两个打靶载体，为该技术的发展奠定了基础。

项目成果

Embryonic stem cells-II : Methods and protocols

胚胎干细胞-II：方法和方案

• 作者: Takahashi M;Haruta M.
• 通讯作者: Haruta M.

ニワトリLIFタンパク質およびその利用

鸡LIF蛋白及其用途

• 发表时间: 2004

Methods in Molecular Biology, vol.329 : Embryonic Stem Cell Protocols

分子生物学方法，第 329 卷：胚胎干细胞方案

• 发表时间: 2005
• 作者: Inada;H.;水野薫;Hiroyuki Horiuchi
• 通讯作者: Hiroyuki Horiuchi

A monoclonal antibody against chicken thrombocytes reacts with the cells of thrombocyte lineage

• DOI: 10.1292/jvms.66.243
• 发表时间: 2004-03-01
• 期刊: JOURNAL OF VETERINARY MEDICAL SCIENCE
• 影响因子: 1.2
• 作者: Horiuchi, H;Tanaka, K;Matsuda, H
• 通讯作者: Matsuda, H

Effect of novel monoclonal antibodies on LIF-induced signal in thicken blastodermal cells

新型单克隆抗体对增厚胚层细胞中 LIF 诱导信号的影响

• 发表时间: 2006
• 期刊: Developmental and Comparative Immunology 30・5
• 作者: Morikawa T;Mizutani M;Aoki N;Watanabe B;Saga H;Saito S;Oikawa A;Suzuki H;Sakurai N;Shibata D;Wadano A;Sakata K;Ohta D;Yusuke Yamashita
• 通讯作者: Yusuke Yamashita