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| BAI1基因敲除打靶载体的快速构建 | |
| 引用本文： | 雷立芳,李大力,周军媚,叶湘漓.BAI1基因敲除打靶载体的快速构建[J].生命科学研究,2012,16(2):95-101. |
| 作者姓名： | 雷立芳 李大力 周军媚 叶湘漓 |
| 作者单位： | 1. 中南大学湘雅三医院神经内科,中国湖南长沙,410013 2. 华东师范大学生命科学学院生命医学研究所,中国上海,200241 3. 湖南师范大学生命科学学院心脏发育研究中心,中国湖南长沙,410081 4. 湖南师范大学生命科学学院心脏发育研究中心,中国湖南长沙410081;湖南师范大学医学院,中国湖南长沙410013 |
| 基金项目： | 湖南省科技厅科技计划项目(2009SK3172);湖南省自然科学基金资助项目(11JJ5071) |
| 摘 要： | BAI1(脑血管生成抑制因子1)因其具有抑制血管生成的作用而得名,研究表明肿瘤的发生可能与BAI1的低表达有关.为了进一步探索BAI1的作用机制,运用改良的Red重组系统和低拷贝中间载体,利用50 bp的同源重组序列直接从BAC载体中克隆长片段的小鼠基因组序列;将得到的基因组序列再次通过重组和改造,构建了BAI1基因的完全敲除并带有报告基因的打靶载体,为后续的构建BAI1基因敲除小鼠模型、在动物体内研究基因功能奠定了基础. |
| 关 键 词： | 基因敲除 打靶载体 Red重组酶 |
Rapid Construction of BAIl Gene-knockout Targeting Vector |
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| LEI Li-fang , LI Da-li , ZHOU Jun-mei , YE Xiang-li.Rapid Construction of BAIl Gene-knockout Targeting Vector[J].Life Science Research,2012,16(2):95-101. | |
| Authors: | LEI Li-fang LI Da-li ZHOU Jun-mei YE Xiang-li |
| Institution: | 3,4(1.The Third Xiangya Hospital,Central South University,Changsha 410013,Hunan,China;2.Institute of Biomedical Sciences,East China Normal University,Shanghai 200241,China;3.College of Life Sciences,Hunan Normal University,Changsha 410081,Hunan,China;4.College of Medicine,Hunan Normal University,Changsha 410013,Hunan,China) |
| Abstract: | Brain angiogenesis inhibitors 1(BAI1) is a transmembrane protein named as its function of inhi-biting the growth of new blood vessels.It has been identified that low expression of BAI1 protein may be associated with the occurrence of tumors.To find out the further function and mechanism of BAI1,a gene knockout mouse will be essential.The first step to generate a gene knockout mouse model is to construct a targeting vector for homologous recombination in mouse embryonic stem cells.Using Red-Cre system as well as Low-and intermediate-copy-number vector,50 bp regions of homology was used to clone long regions of mouse genome from BAC strain.Then the obtained genomic sequence was modified and recombined again,and the aimed complete-knockout targeting vector of BAI1 gene which carried reporter gene was constructed successfully.It has been confirmed that the BAI1 targeting vector is important to its continuing studies of constructing the knockout mouse model and revealing the role of BAI1 gene in animals. |
| Keywords: | gene knocking-out gene targeting vector Red recombinase |
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