中国组织工程研究Issue(7)：1057-1062,6.DOI:10.3969/j.issn.2095-4344.2014.07.013
靶向端粒酶反转录酶基因短发夹RNA载体的构建
Construction of targeting short hairpin RNA plasmid vector expressing TERT gene
摘要
Abstract
BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation. <br> OBJECTIVE:To construct the targeting short hairpin RNA plasmid vector expressing TERT gene from astrocytes by using pLentilox3.7.U6. <br> METHODS:By using two sequences from TERT gene, we synthetized sense and antisense strand template sequences of RNA interference molecular in vitro, and then obtained the complementary strands through annealing procedure. We connected the strands with pLentilox3.7.U6 that was sequenced and transfected into the Escherichia coli. In the end, we tested its effect of reducing the TERT gene expressing by using cultured astrocytes from rat spinal cord in vitro through western blot and immunofluorescence technique. <br> RESULTS AND CONCLUSION:Western blot and immunofluorescence assay showed that, compared with the control group, the interference groups had a lower TERT expression in astrocytes. The targeting short hairpin RNA plasmid vector expressing TERT gene is useful to reduce the TERT gene expression. The targeting short hairpin RNA plasmid vector expressing TERT gene is valid for us to do the further test learning the mechanism of astrocytes in spinal cord injury.关键词
组织构建/组织工程/RNA干扰/端粒酶/端粒酶反转录酶/质粒载体/星形胶质细胞/短发夹RNA/脊髓损伤/胶质瘢痕/国家自然科学基金Key words
transfection/gene expression/cells/plasmids/spinal cord injuries分类
医药卫生引用本文复制引用
宋扬,徐韬,杨明坤,盛伟斌..靶向端粒酶反转录酶基因短发夹RNA载体的构建[J].中国组织工程研究,2014,(7):1057-1062,6.基金项目
国家自然科学基金资助项目(81060106)@@@@the National Natural Science Foundation of China, No.81060106 （81060106）
