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有效PGK1短发夹RNA序列的筛选 被引量：1

Screening of the sequence of short hairpin RNA-specific PGK1 gene
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摘要 目的筛选有效抑制磷酸甘油酸激酶1(PGK1)基因的短发夹核糖核酸(shRNA)序列。方法针对PGK1的不同基因区域设计4个shRNA序列,应用LipofectamineTM2000方法转染至胶质瘤细胞株U251细胞,荧光显微镜下观察shRNA的转染效率。采用RT-PCR方法检测PGK1的mRNA表达,Western blot方法检测PGK1的蛋白表达。结果针对PGK1-homo-441位点的shRNA4(5′-GCAAGGATGTTCTGTTCTTGA-3′)能更有效地抑制PGK1的mRNA及蛋白表达。结论shRNA4是能有效抑制PGK1表达的序列,可用于PGK1的功能研究。 Objective To screen the effective sequence of short hairpin RNA （shRNA） inhibiting PGK1 gene. Methods The shRNAs for PGK1 were transfected to U251 cells using lipofectamineTM 2000. Transfeetion efficiency was confirmed by inverted fluorescent microscopy. RT-PCR and Western blot analysis were performed to detect the expressions of PGK1 mRNA and protein. Results The 4th shRNA in allusion PGKl-homo-441 was more efficient in inhibiting the expressions of PGK1 mRNA and protein. Conclusion The 4th shRNA can efficiently inhibit the production of PGK1, which can be used for studying the effect of PGK1 in the future.
作者 丁浩 成宜军 肖红 阎华 张锐 章文斌 张岩松 邹元杰 刘宏毅
机构地区 南京医科大学附属脑科医院神经外科 南京医科大学附属脑科医院研究所
出处 《江苏医药》 CAS 北大核心 2013年第16期1861-1864,共4页 Jiangsu Medical Journal
基金 国家自然科学基金(NSFC81172390) 南京市医学科技发展项目(ZKX10021)
关键词 短发夹核糖核酸 磷酸甘油酸激酶1基因 Short hairpin RNA PGK1 gene
分类号 R741 [医药卫生—神经病学与精神病学]
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参考文献8

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同被引文献14

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江苏医药
2013年 第16期

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