重组腺病毒表达载体pDC316/PEDF的构建

目的：构建色素上皮衍生因子（PEDF）基因腺病毒表达载体，为老年性黄斑变性基因治疗奠定基础。方法：从大鼠视网膜组织中提取总RNA，RT—PCR扩增PEDF基因eDNA，并将回收的PEDF基因克隆入质粒pGEM—Teasy载体中，亚克隆入腺病毒表达载体质粒pDC316中，并以DNA序列分析加以证实。结果：基因测序结果表明，所克隆的PEDF基因包含了大鼠PEDF基因阅读框内的全部序列，与NCBI Sequence Viewer中公布的大鼠PEDFmRNA序列（NM-177927）完全一致。结论：PEDF基因腺病毒表达载体的成功构建，为下一步体外包装成病毒颗粒研究基因治疗脉络膜新生血管奠定了基础。

Construction of Recombinant Adenovirus Expression Vector pDC316/PEDF

Objective: To construct the adenovirus expression vector of PEDF for gene therapy of age-related macular degeneration (AMD). Method: The PEDF cDNA were amplified by RT-PCR using template mRNA isolated from the tissue of the retina of the rattus. The cDNA fragments were cloned into the vector pGEM-Teasy. The PEDF coding sequence was subcloned into a adenovirus expression plasmid pDC316 and then sequenced. Results: As compared with the sequences of the rat PEDF mRNA published in NCBI Sequence Viewer (NM-177927), the PEDF cDNA fragment contained all the sequences of open reading frame of the rattus PEDF gene. Conclusion: The successful construction of recombination plasmid pDC316/PEDF provides a sound basis for further research on gene therapy of choroidal neovascularization.