脂质体介导质粒pcDNA3.1/ KDRn3基因在转染小鼠视网膜组织中的表达及定位

J4 ›› 2010, Vol. 36 ›› Issue (3): 496-499.

脂质体介导质粒pcDNA3.1/ KDRn3基因在转染小鼠视网膜组织中的表达及定位

左玲¹|栾永昕²|姜扬|裴颖¹|苏冠方¹

1.吉林大学第二医院眼科,吉林长春130041；2.解放军第208医院神经外科|吉林长春130062

收稿日期:2009-12-01 出版日期:2010-05-28 发布日期:2010-05-28
通讯作者: 苏冠方（Tel:0431-88766001，E-mail:lingzuo@sina.com） E-mail:lingzuo@sina.com
作者简介:左玲（1973-）|女|吉林省长春市人|主治医师|医学博士|主要从事眼底病方面的研究。
基金资助:
吉林省科技厅白求恩医学专项基金资助课题（200705231）；吉林省长春市科技计划项目资助课题（2006104）

Expression and location of pcDNA3.1/ KDRn3 gene in retina of mice mediated by liposome transfection

ZUO Ling¹, LUAN Yong-Xi², JIANG Yang, FEI Ying¹, SU Guan-Fang¹

1.Department of Ophthamology,Second Hospital,Jinlin University|Changchun 130041,China;2.Department of |Neurosurgery,NO. 208 Hospital of PLA,Changchun 130062,China

Received:2009-12-01 Online:2010-05-28 Published:2010-05-28

摘要/Abstract

摘要：

目的：将真核表达质粒pcDNA3.1/ KDRn3以脂质体介导转染C57BL/6J小鼠视网膜组织，观察其表达及定位。方法：首先对真核表达质粒pcDNA3.1/KDRn3进行扩增和酶切鉴定，然后将20只健康C57BL/6J小鼠随机分成对照组及玻璃体腔注射后2、5、7和14 d组，每组4只鼠8眼。注射组每眼玻璃体腔注射脂质体pcDNA3.1/ KDRn3复合物1 μL，分别于注射后2、5、7及14 d摘除眼球，制成石蜡切片，用KDRn3蛋白免疫荧光染色在激光共聚焦显微镜下观察KDRn3蛋白表达及定位。结果：Xho Ⅰ和BamHⅠ进行双酶切,切下900 bp大小片段,与目的片段大小一致,表明该质粒确实为KDRn3。注射后2 d在视网膜神经节细胞层细胞胞浆内可见红色荧光信号，注射后5和7 d组视网膜神经节细胞层、内核层部分细胞胞浆内可见红色荧光信号；注射后14 d视网膜神经节细胞层、部分内核层细胞胞浆内可见红色荧光信号，但发出红色荧光信号的细胞数明显减少。　结论：阳离子脂质体可有效将pcDNA3.1/ KDRn3基因转染小鼠视网膜组织，并得到持续表达。

关键词: 血管内皮生长因子受体;基因转染;视网膜

Abstract:

Abstract:Objective To observe the expression and location of the plasmid pcDNA3.1/ KDRn3 after transfected into the retina of C57BL/6J mice by using liposome. Methods The KDRn3 gene was amplified by PCR and then identified by Enzymatic digestion.20 C57BL/6J mice were randomly divided into 5 groups(n＝4),four groups were intravitreally injected with the optimal liposome plasmid mixture,the other was control group. 2,5,7,and 14 d after injection,the localization of pcDNA3.1/ KDRn3 was observed by using immunfluorescence staining method. Results A 900 bp gene part was digested with Xho Ⅰ and BamHⅠwhich agreed with the length of KDRn3.The red fluorescence of KDRn3 protein expressed in the retinal ganglion layer 2 d after injection;after 5 d or 7 d,it expressed both in the ganglion layer and in the inner layer.After 14 d,it showed the weakened expression.Conclusion Cationic liposome can mediate pcDNA3.1/KDRn3 gene into the retina of the C57BL/6J mice effectively and it can express for a long time.

Key words: VEGF receptor;gene transfection;retina

中图分类号:

左玲, 栾永昕, 姜扬, 裴颖, 苏冠方. 脂质体介导质粒pcDNA3.1/ KDRn3基因在转染小鼠视网膜组织中的表达及定位[J]. J4, 2010, 36(3): 496-499.

ZUO Ling, LUAN Yong-Xi, JIANG Yang, FEI Ying, SU Guan-Fang. Expression and location of pcDNA3.1/ KDRn3 gene in retina of mice mediated by liposome transfection[J]. J4, 2010, 36(3): 496-499.