放射诱导调控序列的合成及其辐射诱导特性的研究  

STUDY ON SYNTHETIC PROMOTERS FOR RADIATION-INDUCIBLE GENE THERAPY

作　　者：余东升[1] 黄洪章[1] 潘朝斌[1] 王建广[1] 马健[1] 王安训[1] 

机构地区：[1]中山大学第二附属医院口腔颌面外科,广州510120

出　　处：《口腔颌面外科杂志》2004年第1期1-4,共4页Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金资助项目 ( 3 0 2 714 2 3 );广东省自然科学基金资助项目 ( 2 1865 )

摘　　要：目的　合成放射诱导调控序列并鉴定其辐射诱导特性。方法　利用人工寡核苷酸片段合成含有 6个重复CArG元件的放射诱导调控序列 ,以绿色荧光蛋白 (GFP)作为报告基因转染Tca8113细胞检测其辐射诱导特性。结果　低剂量放射线照射可诱导这种人工调控序列增强GFP在Tca8113细胞中的表达 ,且 3Gy剂量最为明显 ,提高到放射前的 16 1%。结论　人工合成放射诱导调控序列在低剂量放射线照射下能明显增强其下游外源性基因的表达 ,为进一步研究放射 -基因治疗奠定了基础。Objective To synthesize promoters of radiation-mediated gene therapy and testify its radiation-inducible property. Methods The promoters for radiation-mediated gene therapy, containing six CArG elements, were synthesized with man-made complementary oligodenoxyrbi onucleotides. The enhancer regions of the CMV IE gene promoter in pCGFP were replaced by the synthetic promoters to construct the reporter plasmids pE-GFP. Green fluorescence protein (GFP) gene was used as reporter gene to analyse the radiation-inducible property. Results The GFP expression in Tca8113 cells, which were transfected with pE-GFP, was increased by induction of low lose γ-radiation. It was evident that the level of GFP expression observed was dependent on radiation dose, with maximal expression seen following 3Gy irradiation by 161% as compared with control group. Conclusion The synthetic promoters are responsive to low dose of ionizing radiation and the CArG elements can service as a molecular switch to enhance down-stream gene expression. The synthetic promoters are expected to make a valuable contribution to the development of future radiation-responsive vectors for gene therapy.

关 键 词：放射诱导启动子 放射-基因治疗 绿色荧光蛋白 放射诱导调控序列 

分 类 号：R346[医药卫生—基础医学]