【摘要】：以大豆基因組文庫Phytozome公布的大豆Williams82基因組序列為參考,應(yīng)用Primer Premier 5.0軟件設(shè)計(jì)引物,用PCR技術(shù)擴(kuò)增了大豆GmWRI1a基因的啟動(dòng)子序列,構(gòu)建了重組克隆載體pGM-TpGmWRI1a,并通過PCR擴(kuò)增對(duì)陽性克隆進(jìn)行鑒定送測序�？寺～@得GmWRI1a基因啟動(dòng)子序列1 686bp,該啟動(dòng)子序列除含有必需的起始轉(zhuǎn)錄位點(diǎn)、TATA-box、CTTA-box外還包含多個(gè)順式作用元件,如光應(yīng)答元件、赤霉素應(yīng)答元件、表達(dá)分生組織相關(guān)元件、抗旱誘導(dǎo)元件等。同時(shí),構(gòu)建了該啟動(dòng)子植物表達(dá)載體pBI-pGmWRI1a,通過PCR擴(kuò)增、限制性酶切對(duì)陽性克隆進(jìn)行了鑒定,為啟動(dòng)子的功能研究奠定基礎(chǔ)。大豆GmWRI1a基因啟動(dòng)子克隆與序列分析,將為進(jìn)一步研究大豆GmWRI1a基因的表達(dá)調(diào)控及其功能分析提供參考。
[Abstract]:The promoter sequence of soybean GmWRI1a gene was amplified by PCR using the primers designed by Primer Premier 5.0 software and the soybean Williams82 genome sequence published by soybean genomic library Phytozome as reference. The recombinant clone vector pGM-TpGmWRI1a, was constructed and the positive clones were identified and sequenced by PCR amplification. The promoter sequence of GmWRI1a gene, 1686 BP, was cloned and obtained. The promoter sequence contains not only the necessary initial transcription site, but also a number of cis-acting elements, such as photoresponders, gibberellin response elements, and meristem related elements. Drought-resistant induction elements, etc. At the same time, the promoter plant expression vector pBI-pGmWRI1a, was amplified by PCR and the positive clones were identified by restriction endonuclease digestion, which laid a foundation for the functional study of the promoter. Cloning and sequence analysis of soybean GmWRI1a gene promoter will provide reference for further study on the regulation and regulation of soybean GmWRI1a gene expression and functional analysis.
【作者單位】： 東北農(nóng)業(yè)大學(xué)/大豆生物學(xué)教育部重點(diǎn)實(shí)驗(yàn)室/農(nóng)業(yè)部東北大豆生物學(xué)與遺傳育種重點(diǎn)實(shí)驗(yàn)室;
【基金】：營養(yǎng)功能型轉(zhuǎn)基因大豆新品種培育(2016ZX08004-003) 2016年東北農(nóng)業(yè)大學(xué)“學(xué)術(shù)骨干”項(xiàng)目
【分類號(hào)】：S565.1;Q943.2
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本文編號(hào)：2290083