本文選題：GLT-基因　切入點(diǎn)：RNA干擾　出處：《生物技術(shù)通訊》2007年01期 　論文類型：期刊論文

【摘要】：目的:研究特異性siRNA對(duì)大鼠神經(jīng)膠質(zhì)細(xì)胞中GLT-1基因的阻抑效果。方法:根據(jù)GLT-1基因的序列特點(diǎn)和RNAi設(shè)計(jì)原則,設(shè)計(jì)其shRNA的核苷酸片段。退火后將其克隆入pSupressorNeo,構(gòu)建可表達(dá)大鼠GLT-1基因siRNA的重組真核表達(dá)質(zhì)粒pSuppressorNeo-GLT-1;利用脂質(zhì)體法將其轉(zhuǎn)染神經(jīng)膠質(zhì)細(xì)胞后,用RT-PCR、Western印跡及免疫熒光法等方法檢測(cè)轉(zhuǎn)染的神經(jīng)膠質(zhì)細(xì)胞中GLT-1基因的表達(dá)水平。結(jié)果:瞬時(shí)轉(zhuǎn)染的神經(jīng)膠質(zhì)細(xì)胞中GLT-1基因的表達(dá)受到明顯抑制,GLT-1蛋白含量明顯下降。結(jié)論:pSuppressor-Neo-GLT-1質(zhì)粒構(gòu)建成功,瞬時(shí)轉(zhuǎn)染神經(jīng)膠質(zhì)細(xì)胞后可以明顯抑制GLT-1基因的表達(dá)。
[Abstract]:Objective: to study the inhibitory effect of specific siRNA on GLT-1 gene in rat glial cells. Methods: according to the sequence characteristics of GLT-1 gene and the principle of RNAi design, The nucleotide fragment of shRNA was designed and cloned into pSupresor Neo. after annealing, the recombinant eukaryotic expression plasmid pSuppressorNeo-GLT-1 was constructed and transfected into glial cells by liposome method, and the recombinant eukaryotic expression plasmid pSuppressorNeo-GLT-1 was constructed. The expression level of GLT-1 gene in transfected glial cells was detected by RT-PCR Western blot and immunofluorescence. Results: the expression of GLT-1 gene in transient transfected glial cells was significantly inhibited and the content of GLT-1 protein was significantly decreased. Conclusion the plasmid pSuppressor-Neo-GLT-1 was constructed successfully. Transient transfection of glial cells significantly inhibited the expression of GLT-1 gene.
【作者單位】： 第四軍醫(yī)大學(xué)唐都醫(yī)院放射科 解放軍第451醫(yī)院放射科
【分類號(hào)】：Q78

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