發(fā)布時(shí)間：2018-04-25 09:39

  本文選題：廣西巴馬小型豬 + 錨蛋白(ANK)��； 參考：《南方農(nóng)業(yè)學(xué)報(bào)》2017年04期

【摘要】：【目的】克隆廣西巴馬小型豬ANK1基因啟動(dòng)子,確定其活性核心區(qū),為研究ANK1基因啟動(dòng)子與肉質(zhì)性狀的相關(guān)性及構(gòu)建動(dòng)物疾病模型打下基礎(chǔ)�！痉椒ā客ㄟ^(guò)在線軟件對(duì)ANK1基因啟動(dòng)子的轉(zhuǎn)錄因子結(jié)合位點(diǎn)進(jìn)行預(yù)測(cè),根據(jù)轉(zhuǎn)錄因子結(jié)合位點(diǎn)設(shè)計(jì)特異引物擴(kuò)增不同長(zhǎng)度的ANK1基因啟動(dòng)子片段,并利用雙熒光素酶試劑盒檢測(cè)其熒光值,以確定不同ANK1基因啟動(dòng)子片段的活性�！窘Y(jié)果】發(fā)現(xiàn)ANK1基因啟動(dòng)子存在1個(gè)轉(zhuǎn)錄起始位點(diǎn)(TSS)、2個(gè)Cp G島和多個(gè)轉(zhuǎn)錄因子結(jié)合位點(diǎn),并以此作為ANK1基因啟動(dòng)子的分段依據(jù),將其分割為P638、P791、P1113、P1163、P1648、P1694、P1796和P2074等8個(gè)不同長(zhǎng)度的目的片段。成功克隆獲得的8個(gè)ANK1基因啟動(dòng)子片段經(jīng)KpnⅠ和HindⅢ雙酶切、T4真核表達(dá)載體連接、細(xì)胞轉(zhuǎn)染等方法構(gòu)建8個(gè)雙熒光素酶重組報(bào)告基因,雙熒光素酶試劑盒檢測(cè)結(jié)果顯示,廣西巴馬小型豬ANK1基因啟動(dòng)子在P1796片段活性最強(qiáng),與其他片段存在顯著差異(P0.05)�！窘Y(jié)論】成功克隆獲得廣西巴馬小型豬ANK1基因啟動(dòng)子的8個(gè)片段,且利用雙熒光素酶試劑盒檢測(cè)確定其核心啟動(dòng)子區(qū)域出現(xiàn)在P1796片段。
[Abstract]:[objective] to clone the promoter of ANK1 gene from Guangxi Bama miniature pig and determine its active core region. In order to study the correlation between ANK1 gene promoter and fleshy traits and to construct animal disease model, the transcription factor binding sites of ANK1 gene promoter were predicted by online software. According to the transcription factor binding site, specific primers were designed to amplify the promoter fragments of ANK1 gene of different lengths, and the fluorescence values of the promoter fragments were detected by double luciferase kit. In order to determine the activity of different promoter fragments of ANK1 gene. [results] it was found that there were one transcriptional initiation site (TSS), two CP G islands and multiple transcription factor binding sites in the promoter of ANK1 gene, which were used as the segmental basis of the promoter of ANK1 gene. It was divided into P638P791P1113P1163, P1648P1694, P1796 and P2074, etc. Eight ANK1 promoter fragments were successfully cloned and ligated into Kpn 鈪，

本文編號(hào)：1800804