系統情報  NBRP Rat No: 0639  系統名: SD-Tg(Gfap-Tk)Jog  通称: GFAP-TK Rat Genome Database 研究責任者:  Jonathan flint          University of Oxford         〒 OX3 7BN   Oxford Roosevelt Drive        Tel: +44 (0)1865 287772     Fax: +44 (0)1865 287501 Email:  jf@well.ox.ac.uk 保存状況：   胚            精子           生体 毛色  White 近交世代数  More than 10 利用条件  利用者は、研究成果の公表にあたって謝辞の表明を必要とする。 分類   Inbred  Segregating   Congenic   Consomic   Recombinant   Coisogenic   Spont. Mutant  Transgene  Ind. Mutant  その他  コマーシャルブリーダー 研究分野   糖尿病・肥満  脳神経疾患  眼疾患   歯科疾患  循環器系疾患・高血圧  がん・腫瘍  代謝、内分泌  耳鼻疾患  免疫・アレルギー疾患  感染症  骨形態異常   消化器疾患  皮膚疾患   生殖器疾患   発生  行動・学習  血液疾患  泌尿器疾患   薬理学  その他    対照動物  レポーター遺伝子動物 遺伝子 Tk: thymidine kinase Gfap: glial fibrillary acidic protein 由来 The rats were originally generated by the University of Michigan Transgenic Animal Model Core, University of Michigan. They use a Crl:CD(SD) sub strain of Sprague-Dawley rats, obtained from Charles River Laboratory (Wilmington, MA, USA). The F1 rats have been bred in the UK with Hsd:Sprague Dawley (Harlan laboratories, UK), which are direct descendants of the original 1925 SD-company colony (Madison, Wisconsin, USA). The GFAP-TK rat was generated by pronuclear injection of a‘Gfap-Tk' Bacterial Artificial Chromosome (BAC) construct, engineered using ‘RedET’-mediated homologous recombination methods (Groves et al., 2013). 特性など Rats show no phenotype until dosed with the antiviral agant ganciclovir. Chronic dosing at 10mg/kg/day via subcutaneous minipump for 21 days results in a loss of DCX-positive cells in the dentate gyrus and subventricular zone. 繁殖・維持方法 Males are infertile. Females generate good litters of 10-15 pups showing 50% heterozygous offspring. 遺伝診断法 GFAP-TK transgene can be identified using the foolwoing primers and protocols: GFAP-TK_genoF cca acg gcg acc tgt ata ac GFAP-TK_genoR gcc ctt tag tga ggg tta att c Reagent Volume (ul) Template DNA (10ng/ul) 2 Forward primer (10mMol) 1 Reverse Primer (10mMol) 1 Buffer (10x) 2 dNTP (8mMol) 2 Mg2+ (25mMol) 1.6 Taq Polymerase (Hot Star, Qiagen) 0.1 ddH20 10.3 Total Volume 20 Step Temp (°C) Time DNA activity 1 98 15 minutes Activation of polymerase 2 95 30s Denaturation 3 64 30s Annealing 4 -0.5 °C per cycle 5 72 1 min Extension 6 Go to step 3, 13 times 7 57 30s Annealing 9 72 1 min Extension 10 Got to step 7, 30 times 11 72 10 min final extension 12 4 forever 参考文献 Groves JO, Leslie I, Huang GJ, McHugh SB, Taylor A, Mott R, Munafò M, Bannerman DM, Flint J. Ablating Adult Neurogenesis in the Rat Has No Effect on Spatial Processing: Evidence from a Novel Pharmacogenetic Model. PLoS Genet. 2013;9(9):e1003718. doi: 10.1371/journal.pgen.1003718. Epub 2013 Sep 5. 系統情報