Chromosome engineering for the development of transgenic avian

用于开发转基因禽类的染色体工程

• 批准号: 11450313
• 负责人: IIJIMA Shinji
• 金额: $ 9.54万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11450313/
• 关键词: Transgenic; PGC; Chicken; Transfection; Recombinant DNA; Integrase; Integration; ヒト成長ホルモン; オボアルブミン

We are trying to establish transgenic chicken by using a virus vector. For this, a pantropic retrovirus which is based on Moloney murine leukemia virus and contains VSV-G gene instead of env. This virus has several advantages such as broad host range. GFP and Neo^r gene. The former gene was expressed from virus LTR and the latter from inserted RSV promoters. A helper cell line which express virus gag and pot genes, was transfected with this vector and stable transfectants were isolated. VSV-G gene was transiently infected to one of the transfectant and replication-deficient virus particles were produced. By ultracentrifigation, virus particles were concentrated to 10^9 cfu/ml. Blastderm of quail was infected with this virus. After in vitro hatching, we analyzed the presence of vector DNA sequences. In almost all cells of Go chimera, this vector sequence could be detected.

我们正在尝试用病毒载体建立转基因鸡。为此，基于Moloney鼠白血病病毒并含有VSV-G基因而不是env的泛嗜性逆转录病毒。该病毒具有宿主范围广等优点。GFP和Neo^r基因。前者基因由病毒LTR表达，后者由插入的RSV启动子表达。用该载体转染表达病毒gag和pot基因的辅助细胞系，并分离稳定的转染子。将VSV-G基因瞬时感染其中一个转染子，产生复制缺陷型病毒颗粒。通过超离心，将病毒颗粒浓缩至10^9 cfu/ml。鹌鹑胚囊感染了该病毒。在体外孵化后，我们分析了载体DNA序列的存在。在Go嵌合体的几乎所有细胞中，都可以检测到该载体序列。

项目成果

Shinji Mizuarai, Masamichi Kamihira, Ken-Ichi Nishijima and Shinji Iijima: "Integrase-mediated nonviral gene transfection with enhanced integration efficiency"Journal of Bioscience and Bioengineering. 88[5]. 461-467 (1999)

Shinji Mizuarai、Masamichi Kamihira、Ken-Ichi Nishijima 和 Shinji Iijima：“整合酶介导的非病毒基因转染，具有增强的整合效率”生物科学与生物工程杂志。

Yuichi Machida,Katsuhide Miyake,Kouji Hattori,Shin Yamamoto,Mitsuo Kawase and Shinji Iijima: "Structure and function of a novel coliphage-associated sialidase"FEMS Microbiology Letters. 182. 333-337 (2000)

Yuichi Machida、Katsuhide Miyake、Kouji Hattori、Shin Yamamoto、Mitsuo Kawase 和 Shinji Iijima：“新型大肠杆菌噬菌体相关唾液酸酶的结构和功能”FEMS 微生物学快报。

Shinji Mizuarai, Masamichi Kamihira, Ken-Ichi Nishijima and Shinji Iijima: "Integrase-mediated nonvital gene transfection with enhanced integration efficiency"Joumal of Bioscience and Bioengineering. 88[5]. 461-467 (1999)

Shinji Mizuarai、Masamichi Kamihira、Ken-Ichi Nishijima 和 Shinji Iijima：“整合酶介导的非生命基因转染，具有增强的整合效率”生物科学与生物工程杂志。