Establishment and analyses of mouse models of glycine-encephalopathy by conditional knockout of mouse H protein-gene.

通过条件性敲除小鼠H蛋白基因建立小鼠甘氨酸脑病模型并进行分析。

基本信息

批准号：
14370052
负责人：
KATO Ichiro
金额：
$ 8.77万
依托单位：
TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370052/
关键词：
glycine H protein ES cells Cre recombinase knockout mouse prenatal death encephalopathy Cre レコンビナーゼ

项目摘要

1.We made the knock-out vector (H protein gene-KO) that carry mouse H protein 5'-genomic DNA, loxP, exon1, loxP, pGK-neo, loxP, mouse H protein 3'-genomic DNA in this sequence.2.We introduced the linearized knock-out vector into mouse ES cells and got 11 cell lines of ES cells that have undergone homologous recombination between the ES genome and knock-out vector.3.Next, we microinjected recombinated ES cells to C57 Black mouse blastocysts and obtained 7 male chimeras, 3 of them have undergone germ-line transmission of the mutated allele.4.We then mated F1 mice with Cre recombinase-expressing transgenic mice. In the next generation, excision of exon1 DNA sequence was confirmed by genome PCR and Southern blot analyses.5.To determine whether the H protein expression is downregulated in the tissues of H protein +/-mice, we performed Western blot analysis using anti-H protein rabbit polyclonal antibody. Western blot analysis indicated that the H protein is decreased to 50% in the brains of +/-mice as compared to +/+ mice.6.We mated males and females of H protein +/-mice and analyzed the genotypes of newborn pups and embryos. PCR analyses have clearly shown that homozygous H protein knockout mice can not be born alive or even develop to E14.7.In the present study, we obtained the H protein +/-mice that have reduced (50%) levels of H protein and showed that H protein is essential for normal development and survival of mouse embryos. By using Cre/lox P system, we will be able to know the unknown functions of H protein in vivo.

1.我们制作了载体载体（H蛋白基因-KO），该载体载有小鼠H蛋白5'基因组DNA，LOXP，EXON1，LOXP，PGK-NEO，LOXP，LOXP，小鼠H蛋白3'-基因组DNA在此序列中。矢量。3.next，我们将重新注射的ES细胞重组为C57黑小鼠胚泡并获得了7个雄性嵌合体，其中3个经历了突变等位基因的细菌线传播。4。然后，我们与CRE重组酶表达的转基因小鼠配对F1小鼠。在下一代中，通过基因组PCR和Southern印迹分析证实了Exon1 DNA序列的切除。5。为了确定H蛋白在H蛋白+/-小鼠的组织中是否将H蛋白表达下调，我们使用抗H蛋白质兔多克隆抗体进行了蛋白质印迹分析。蛋白质印迹分析表明，与 +/ +小鼠相比，+/-小鼠的大脑中的H蛋白降至50％。6。我们配对H蛋白+/-小鼠的男性和女性，并分析了新生幼崽和胚胎的基因型。 PCR分析清楚地表明，纯合H蛋白基因敲除小鼠不能活着，甚至无法发育为E14.7。在本研究中，我们获得了H蛋白+/-小鼠的H蛋白（50％）H蛋白水平，并表明H蛋白对于小鼠胚胎的正常发育和存活至关重要。通过使用CRE/LOX P系统，我们将能够了解体内H蛋白的未知功能。

项目成果

期刊论文数量（79）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Eifuku S.: "Neuronal correlates of face identification in the monkey anterior temporal cortical areas"J.Neurophysiol.. 91・1. 358-371 (2004)

Eifuku S.：“猴子前颞皮质区域面部识别的神经相关性”J.Neurophysiol.. 91・1（2004）。

DOI：
发表时间：
期刊：
影响因子：
0
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Comparison of brain activity between dopamine d2 receptor-knockout and wild mice in response to dopamine agonist and antagonist assessed by FMRI.

通过 FMRI 评估多巴胺 d2 受体敲除小鼠和野生小鼠对多巴胺激动剂和拮抗剂反应的大脑活动的比较。

DOI：
发表时间：
2004
期刊：
NeuroSignals 3
影响因子：
0
作者：
Kuriwaki;J.;Nishijo;H.;Kondoh;T.;Uawano;T.;Torii;K.;Katsuki;M.;Ono T.
通讯作者：
Ono T.

Deficit of CD38/cyclic ADP-ribose is differentially compensated in hearts by gender.

DOI：
10.1016/j.bbrc.2003.10.143
发表时间：
2003-12
期刊：
Biochemical and biophysical research communications
影响因子：
3.1
作者：
J. Takahashi;Y. Kagaya;I. Kato;J. Ohta;S. Isoyama;M. Miura;Y. Sugai;M. Hirose;Y. Wakayama;M. Ninomiya;J. Watanabe;S. Takasawa;H. Okamoto;K. Shirato
通讯作者：
J. Takahashi;Y. Kagaya;I. Kato;J. Ohta;S. Isoyama;M. Miura;Y. Sugai;M. Hirose;Y. Wakayama;M. Ninomiya;J. Watanabe;S. Takasawa;H. Okamoto;K. Shirato

Fukuda Y.: "Regulated transgene delivery by ganciclovir in the brain without physiological alterations by a live attenuated herpes simplex virus vector"Neurosci.Res.. 45・2. 233-241 (2003)

Fukuda Y.：“通过活的减毒单纯疱疹病毒载体在大脑中调节更昔洛韦的转基因递送，而没有生理改变” Neurosci.Res.. 233-241 (2003)。