视网膜特异性Rho启动子的基因克隆与序列分析  

Gene Clone and Sequence Analysis of Retinal Specific Rho Promoter

作　　者：吕秀兰[1] 吕林[2] 李永浩[2] 张静琳[2] 李石毅[2] 

机构地区：[1]广州市番禺区人民医院眼科,广州511400 [2]中山大学中山眼科中心,广州510060

出　　处：《眼科学报》2005年第2期119-121,共3页Eye Science

基　　金：广州市科技局基金资助项目(A052001084)

摘　　要：目的:获取视网膜特异性表达Rho启动子基因,为实现外源基因在视网膜组织中特异性表达做准备。方法:用酚氯仿异戊醇抽提BLAB/c鼠全基因组DNA;设计引物,从基因组中通过PCR扩增获得视网膜特异性表达的Rho启动子基因;克隆入PcDNA3.1+载体,酶切、PCR鉴定,上海博亚生物技术公司测序;序列分析。结果:从BLAB/c鼠全基因组DNA中PCR扩增得到预期大小的启动子片段;构建重组PcDNA3.1+-Rho载体具有与目标片段长度相符的插入片段,插入方向正确;测序结果分析显示,视网膜特异性Rho启动子高度保守,所获得的BLAB/c鼠Rho启动子与Genebank报道的序列一致。结论:成功获得BLAB/c鼠Rho视网膜特异性启动子,为下一步研究视网膜疾病的发病机制和基因治疗奠定了一定的工作基础。Purpose:To obtain the gene encoding retinal specific Rho promoter therefore to prepare for exogenous gene transcription and expression in retina. Methods:Extract the genomic DNA of a BLAB/c mice .The Rho promoter DNA were then amplified by PCR,the product of which was subcloned into PcDNA3.1+vector. After identification by restriction enzymes, the recombinant PcDNA3.1+vectors were subjected to sequence analysis. Results:The fragments of the promoter as amplified by PCR were of predicted length. Digestion with HindIII and NheI proved correct insertion of the target fragments with expected length into the recombinant PcDNA3.1+vectors. As indicated by homology analysis the Rho promoter was highly conservative, and the Rho promoter we got had hole homology as reported by Boulanger A etc. Conclusion:Retinal specific Rho promoter has been successfully cloned,thus making possible the subsequent research as the mechanism and gene therapy of retina.

关 键 词：克隆与序列分析 RHO BLAB/c鼠 基因组DNA PCDNA3 特异性表达 启动子基因 PCR扩增 生物技术公司 视网膜组织 PCR鉴定 视网膜疾病 外源基因 设计引物 插入片段 片段长度 基因治疗 发病机制 异戊醇 克隆人 步研究 载体 测序 

分 类 号：R774.1[医药卫生—眼科]