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| BPOZ基因剔除小鼠模型的建立 | |
| 引用本文： | 项佑贵,孙霞,王龙,严兰珍,杨桦,刘伟,许勇,徐国江,王一,费俭,傅继梁,王铸钢.BPOZ基因剔除小鼠模型的建立[J].生物化学与生物物理进展,2004,31(7):616-621. |
| 作者姓名： | 项佑贵 孙霞 王龙 严兰珍 杨桦 刘伟 许勇 徐国江 王一 费俭 傅继梁 王铸钢 |
| 作者单位： | 1. 上海第二医科大学基础医学院医学遗传学教研室,上海,200025;中国科学院上海生命科学研究院-上海第二医科大学健康科学中心,上海,200025 2. 上海南方模式生物研究中心,上海,201203 3. 上海南方模式生物研究中心,上海,201203;上海第二医科大学附属瑞金医院医学基因组学国家重点实验室,上海,200025;中国科学院上海生命科学研究院-上海第二医科大学健康科学中心,上海,200025 4. 上海第二医科大学基础医学院医学遗传学教研室,上海,200025;上海南方模式生物研究中心,上海,201203;上海第二医科大学附属瑞金医院医学基因组学国家重点实验室,上海,200025;中国科学院上海生命科学研究院-上海第二医科大学健康科学中心,上海,200025 |
| 基金项目： | 国家自然科学基金资助项目（30370793），国家杰出青年科学基金（39925023），教育部“长江学者奖励计划”（OOTPJS111），国家高技术“863”计划(2001AA216081)，上海市科学技术发展基金（99JC14029,99XD14005）资助项目. |
| 摘 要： | BPOZ是在卵巢癌等肿瘤组织中表达下调的细胞生长抑制基因，建立BPOZ基因剔除小鼠模型，可以为在体研究BPOZ基因的生物学功能及其与肿瘤发生的关系创造条件.运用生物信息学手段确定小鼠BPOZ基因组序列,设计基因剔除策略，构建完成了基因剔除载体XpPNT-BPOZ.以电穿孔方法将基因剔除载体导入ES细胞，用G418和Ganciclovoir进行正负筛选，获得抵抗克隆，PCR和DNA印迹鉴定出正确同源重组的ES细胞克隆.将同源重组的ES细胞注入小鼠囊胚，获得嵌合体小鼠.嵌合体小鼠与C57BL/6J小鼠交配后获得Aguoti毛色的小鼠30只，其中15只为BPOZ基因剔除杂合子小鼠，阳性率为50%.在雌雄杂合子交配的后代中获得纯合子小鼠.初步的表型观察发现BPOZ基因剔除小鼠发育正常，有繁殖能力，进一步的表型分析工作正在进行之中. |
| 关 键 词： | BPOZ基因，细胞生长抑制基因，基因剔除 |
| 收稿时间： | 1/7/2004 12:00:00 AM |
| 修稿时间： | 2/6/2004 12:00:00 AM |
Generation of BPOZ Gene Knock-out Mouse Model |
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| XIANG You-Gui,SUN Xi,WANG Long,YAN Lan-Zhen,YANG Hu,LIU Wei,XU Yong,XU Guo-Jiang,WANG Yi,FEI Jian,FU Ji-Liang and WANG Zhu-Gang.Generation of BPOZ Gene Knock-out Mouse Model[J].Progress In Biochemistry and Biophysics,2004,31(7):616-621. | |
| Authors: | XIANG You-Gui SUN Xi WANG Long YAN Lan-Zhen YANG Hu LIU Wei XU Yong XU Guo-Jiang WANG Yi FEI Jian FU Ji-Liang and WANG Zhu-Gang |
| Institution: | Laboratory of Medical & Molecular Genetics, Department of Medical Genetics, Shanghai Second Medical University, Shanghai 200025, China; Health Science Center of Shanghai Institute of Biological Science of The Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China; State Key Laboratory for Medical Genomics, Rui-Jin Hospital Affiliated to Shanghai Second Medical University, Shanghai 200025, China;Health Science Center of Shanghai Institute of Biological Science of The Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Laboratory of Medical & Molecular Genetics, Department of Medical Genetics, Shanghai Second Medical University, Shanghai 200025, China; Health Science Center of Shanghai Institute of Biological Science of The Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Shanghai Research Center for Model Organisms, Shanghai 201203, China;Laboratory of Medical & Molecular Genetics, Department of Medical Genetics, Shanghai Second Medical University, Shanghai 200025, China; Shanghai Research Center for Model Organisms, Shanghai 201203, China; State Key Laboratory for Medical Genomics, Rui-Jin Hospital Affiliated to Shanghai Second Medical University, Shanghai 200025, China;Health Science Center of Shanghai Institute of Biological Science of The Chinese Academy of Sciences and Shanghai Second Medical University, Shanghai 200025, China |
| Abstract: | BPOZ is one protein containing ankyrin repeat and BTB (POZ) domain which gene is located at 3q21.3 in human genome. It was found that the expression of BPOZ gene is down-regulated in human ovary tumors, suggesting it is potentially a cell growth or tumor suppressor gene. BPOZ gene knockout mouse model was established for further in vivo study of its normal function and the role of BPOZ in tumorigenesis. The mouse genomic DNA sequence of BPOZ gene was verified through bioinformatic analysis. According to the BPOZ genomic DNA sequence, the strategy of gene targeting and constructing of knockout vector (XpPNT-BPOZ) were established to delete the region of mouse genome, which spans from exon 2 to 12 of BPOZ gene. The gene knockout vector XpPNT-BPOZ was constructed and confirmed by restriction enzyme digestion and sequencing. Electroporation of ES cells with XpPNT-BPOZ and screening of both G418 and Ganciclovoir resistant clones were performed according to common protocol. The homologous recombined ES cell clones were identified by PCR and confirmed by Southern blot analysis. After transplantation of homologous recombined ES cells into blastocysts through microinjection and mating of chimeras with C57BL/6J mice, 30 offspring with Aguoti fur in color were acquired. 15 of them (50%) show genotype heterozygous for BPOZ. The BPOZ heterozygotes were intercrossed to generate mutants homozygous. Finally, the cohort of mutants homozygous for BPOZ was established. The mutants are fertile, and further phenotype analysis is under way. |
| Keywords: | BPOZ gene cell growth suppressor gene gene knockout |
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