SNP标记对角膜混浊小鼠突变相关基因的精细定位-遗传.PDFVIP

HEREDITAS (Beijing) 2010 5 , 32(5): 486―491 ISSN 0253-9772 研究报告 DOI: 10.3724/SP.J.1005.2010.00486 SNP 标记对角膜混浊小鼠突变相关基因的精细定位 1 2 1 1,2 蒋荧梅 , 刘春 , 吴刘成 , 邵义祥 1. , 226001; 2. , 226001 为深入研究前期工作中以 ENU 诱变技术建立的遗传性角膜混浊突变系小鼠(B6-Co) 的遗传机制, 利用 SNP 标记对其突变基因进行精细定位, 将该品系中具有角膜混浊表型的小鼠(B6-CoP)与 DBA/2 小鼠(简称 D2) 配种得到F1 代, 再回交D2 亲本品系得到F2 代, 提取F2 代角膜混浊小鼠鼠尾DNA 。在MGI 数据库中选取小鼠 13 号染色体已定位区间附近5 个在 C57BL/6(简称B6)和D2 两个品系之间有差异的 SNP, 应用聚合酶链反应- 限制性片段长度多态性(PCR-RFLP)技术及连锁分析方法对 B6-Co 小鼠突变基因进行精细定位。结果表明: B6-Co 小鼠突变基因定位于 13 号染色体上 112 546 283~113 397 654 bp 之间, 因该区间内有5 个已知基因, 其 中Map3k1 基因与小鼠眼睛形态生成和眼睑闭合密切相关, 提示Map3k1 是B6-Co 小鼠突变的强力候选基因。 SNP; 角膜混浊小鼠; 突变基因; 基因定位 Fine mapping of mutant gene related corneal opacity mouse with SNPs 1 2 1 1, 2 JIANG Ying-Mei , LIU Chun , WU Liu-Cheng , SHAO Yi-Xiang 1. Laboratory Animals Centre , Nantong University, Nantong 226001, China; 2. Institute of Comparative Medicine, Nantong University, Nantong 226001, China Abstract: To further investigate the genetic mechanism of the mutant mice(B6-Co) with hereditary corneal opacity phe- notype obtained by ENU-induced mutagenesis from B6 in previous study, SNP markers were used to map the mutant gene of B6-Co mice. F generation mice were bred by backcrossing (B6-CoP×D2 )F with D2 and the DNA samples of F mutant 2 1 2 mice were extracted from the tails. Five SNP sites that showed differences between B6 and D2 strains nearby the located region on chromosome 13 were screened from MGI database. Five SNPs, PCR-RFLP and linkage analyses were carried out to map the