【摘 要】
：
目的：病理性近视是目前最常见的难治性致盲眼病之一,人Lumican基因(LUM基因)是重要候选基因,建立携带突变位点(cDNA596T＞C)的LUM基因的转基因鼠模型,为深入研究病理性近视的发病机制奠定基础.方法：利用基因重组技术,将人工合成的LUM基因第二外显子596位碱基T替换为C,再克隆到质粒pRPDes3d的多克隆位点,构建pRP.E×3d-EF1A＞LUM/flag＞IRES/hrGFP
【机 构】
：
首都医科大学附属北京同仁医院,北京同仁眼科中心,北京市眼科学与视觉科学重点实验室,北京,100730
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目的：病理性近视是目前最常见的难治性致盲眼病之一,人Lumican基因(LUM基因)是重要候选基因,建立携带突变位点(cDNA596T＞C)的LUM基因的转基因鼠模型,为深入研究病理性近视的发病机制奠定基础.方法：利用基因重组技术,将人工合成的LUM基因第二外显子596位碱基T替换为C,再克隆到质粒pRPDes3d的多克隆位点,构建pRP.E×3d-EF1A＞LUM/flag＞IRES/hrGFP质粒载体；然后用显微注射法将其转至BDF1小鼠受精卵雄原核中,存活的受精卯再移入ICR假孕雌鼠输卵管内,得到可表达LUM基因的F0代转基因小鼠C57-TgN(LUM)CCMU.经PCR、Westem印迹杂交法分析子代鼠中基因整合及表达的情况.结果：得到31只新生仔鼠,PCR检测有6只LUM阳性转基因鼠,western印迹杂交法证明转基因阳性鼠转入的目的基因片段确实合成了相应蛋白质.稳定遗传3代,共获得128只仔鼠,经PCR检测其中有68只为LUM阳性,阳性率达53.13％.结论：携带突变位点(cDNA 596T＞C)的LUM基因的转基因鼠模型构建成功,为进一步探索LUM基因突变对病理性近视的发生发展及细胞外基质成分的影响提供了重要的研究平台,为病理性近视发病机制的研究奠定了基础.
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