Abstract: In order to create an N‑ethyl‑N‑nitrosourea（ENU）‑induced eye disease mouse model，C57BL/6J male mice were injected intraperitoneally with ENU，HE and immunohistochemical staining analysis of eye sections were conducted for eye malformed mouse，and linkage and sequence analysis were conducted to determine the mutant gene.HE staining analysis showed vacuoles in the surface of the corneal epithelial cells，collagen fibrils were loose in the corneal stromal layer and some endothelial cells were absent in mutant corneas. Immunohistochemistry showed abnormal corneal epithelium differentiation in mutant corneas. The mutation was mapped to mouse chromosome 2 between markers D2Mit107 and D2Mit423，which were located at 65.13 and 73.57 cM，respectively.The Jag1 gene was finally selected as a candidate gene for this mutation.Automated sequencing revealed that the AG base at 3′ end of the intron 24 in the Jag1 gene on one chromosome of the mutant heterozygous mouse was replaced by the GG base，resulting in the deletion of 7 bases in the exon 25.Translation of the mutant allele would produce frameshift and truncated protein.The results showed that the Jag1 gene mutation was associated with the eye malformation phenotype.

Key words: Mouse, N?ethyl?N?nitrosourea（ENU）, Eye malformation, Gene mapping, Jag1 gene

摘要： 为建立人类和动物眼病的小鼠模型，采用乙烷基亚硝基脲（ENU）诱导C57BL/6J（B6）小鼠突变手段获得眼病小鼠，利用苏木精-伊红（HE）染色法及免疫组织化学染色技术分析小鼠角膜病变特点，通过连锁分析及位置候选克隆确定致病基因。结果显示，ENU诱变获得1例眼畸形小鼠，小鼠角膜上皮细胞层出现明显空泡，且分化异常，角膜基质层胶原纤维排列杂乱松散，角膜内皮细胞部分脱落。染色体定位将致病基因定位于小鼠第2号染色体微卫星D2Mit107（距着丝粒65.13 cM）与D2Mit423（距着丝粒73.57 cM）之间。对候选基因齿状基因1（Jag1）测序发现，突变杂合子小鼠1条染色体上Jag1基因第24号内含子3′端AG碱基被GG碱基替代，导致第25号外显子存在7个碱基的缺失；蛋白质预测发现，该突变引起Jag1 编码区移码及蛋白质编码提前终止。综合分析可知，Jag1基因突变是引起小鼠眼畸形表型的分子基础。

关键词: 小鼠, 乙烷基亚硝基脲, 眼畸形, 基因定位, 齿状基因1