本文選題：人Tudor-SN蛋白 + ��； 參考：《山東醫(yī)藥》2017年04期

【摘要】：目的為深入研究Tudor-SN蛋白本身在翻譯水平的調(diào)控機制奠定基礎(chǔ)。方法以HeLa細(xì)胞全基因組DNA為模板,PCR擴增出目的基因,利用雙酶切的方法將目的片段連接到pGL3-Control載體上。再將構(gòu)建成功的pGL3-5'UTR、pGL3-3'UTR重組質(zhì)粒分別與內(nèi)參海腎熒光素酶質(zhì)粒瞬時共轉(zhuǎn)染HeLa細(xì)胞,培養(yǎng)48h檢測熒光素酶的活性。結(jié)果雙酶切和基因測序法鑒定重組質(zhì)粒構(gòu)建成功,轉(zhuǎn)染重組質(zhì)粒后可檢測到熒光素酶的活性;以pGL3-5'UTR質(zhì)粒熒光素酶活性最高。結(jié)論成功構(gòu)建了人Tudor-SN基因5'UTR和3'UTR序列的重組質(zhì)粒,為研究Tudor-SN基因UTR區(qū)對翻譯調(diào)控的影響奠定了基礎(chǔ)。
[Abstract]:Objective to lay a foundation for further study on the regulation mechanism of Tudor-SN protein in translation level. Methods the target gene was amplified from the whole genomic DNA of HeLa cells and ligated to the pGL3-Control vector by double enzyme digestion. Then the constructed recombinant plasmid pGL3-5 UTR-pGL3-3 UTR was cotransfected into HeLa cells at the same time, and the luciferase activity was detected after 48 h culture. Results the recombinant plasmid was successfully constructed by double enzyme digestion and gene sequencing. The luciferase activity of the recombinant plasmid was detected after transfection, and the luciferase activity of the pGL3-5'UTR plasmid was the highest. Conclusion Recombinant plasmids of 5'UTR and 3'UTR of human Tudor-SN gene were successfully constructed, which laid a foundation for studying the effect of UTR region of Tudor-SN gene on translation regulation.
【作者單位】： 天津醫(yī)科大學(xué);
【基金】：國家杰出青年基金資助項目(31125012) 教育部“創(chuàng)新團隊發(fā)展計劃”(IRT13085) 國家自然科學(xué)基金資助項目(31170830/31370749/31571380) 天津市應(yīng)用基礎(chǔ)與前沿技術(shù)研究計劃(青年基金項目)(15JCQNJC09900) 天津醫(yī)科大學(xué)青年基金項目(2015KYZQ03)
【分類號】：Q78
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本文編號：1843932