本文選題：pRFP　切入點(diǎn)：啟動(dòng)子　出處：《廣東醫(yī)學(xué)》2017年02期

【摘要】：目的構(gòu)建人p53RFP基因啟動(dòng)子序列不同截短片段的熒光素酶報(bào)告基因載體,研究啟動(dòng)子的轉(zhuǎn)錄活性。方法 PCR擴(kuò)增人p53RFP基因啟動(dòng)子區(qū)域不同長(zhǎng)度的目的片段,克隆到pGL3-Basic中,構(gòu)建啟動(dòng)子區(qū)域3個(gè)不同截短片段的熒光素酶報(bào)告基因載體,經(jīng)雙酶切和基因測(cè)序鑒定正確后,轉(zhuǎn)染HEK293細(xì)胞,雙熒光素酶報(bào)告基因檢測(cè)系統(tǒng)檢測(cè)熒光素酶活性。結(jié)果成功構(gòu)建了p53RFP啟動(dòng)子不同截短片段的熒光素酶報(bào)告基因載體,經(jīng)雙熒光素酶報(bào)告基因檢測(cè)分析,3個(gè)啟動(dòng)子片段均具有轉(zhuǎn)錄活性。結(jié)論成功構(gòu)建了人p53RFP基因啟動(dòng)子熒光素酶報(bào)告基因載體,為研究p53RFP基因的轉(zhuǎn)錄調(diào)控機(jī)制提供了實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:Objective to construct a luciferase reporter gene vector with different truncated fragments of human p53RFP promoter sequence and to study the transcriptional activity of promoter. Methods the target fragments with different length in the promoter region of human p53RFP gene were amplified by PCR and cloned into pGL3-Basic. Three luciferase reporter gene vectors with different truncated fragments in promoter region were constructed and identified by double enzyme digestion and gene sequencing, and then transfected into HEK293 cells. The luciferase activity was detected by double luciferase reporter gene detection system. Results the luciferase reporter gene vector with different truncated fragments of p53RFP promoter was constructed successfully. Conclusion Human p53RFP gene promoter luciferase reporter gene vector was constructed successfully, which provides an experimental basis for studying the transcriptional regulation mechanism of p53RFP gene.
【作者單位】： 廣州醫(yī)科大學(xué)附屬第二醫(yī)院心內(nèi)科廣州心血管疾病研究所;南方醫(yī)科大學(xué)南方醫(yī)院心內(nèi)科;廣州軍區(qū)廣州總醫(yī)院干部病房二科;
【基金】：國(guó)家自然科學(xué)基金青年基金資助項(xiàng)目(編號(hào):30800462) 廣州醫(yī)科大學(xué)博士啟動(dòng)項(xiàng)目(編號(hào):2012C45)
【分類號(hào)】：R3416

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