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求翻譯一段文字 Assessment of the stability of chlorinated alkaloid production in subsequent subcultures. Ten root tips from hairy roots transformed with pCAMRebH/RebF and pCAMPyrH/RebF/STRvm were subcultured every 3 weeks in Gamborg’s B5 solid medium (half-strength basal salts, full-strength vitamins, 30 g l21 sucrose, 6g l21 agar, pH 5.7). and grown at 26 uC in the dark. Alkaloids were isolated from 21-day-old hairy roots and analysed as described above (Supplementary Figs 11–14). Verification of expression of RebH, RebF and PyrH, STRvm enzymes by realtime RT–PCR. Real-time RT–PCR was used to assess the expression levels of RebH and RebF. Expression levels in hairy roots infected with A. rhizogenes lacking the pCAMRebH/RebF construct were compared with expression levels in hairy roots harbouring pCAMRebH/RebF. Messenger RNA from transformed hairy roots was isolated and purified from contaminant DNA using a Qiagen RNeasy Plant Mini Kit and Rnase-free DnaseI, respectively. The resulting mRNA was then reverse-transcribed to cDNA using a Qiagen QuantiTect Reverse transcription kit and then subjected to PCR with specific primers (see below), a Qiagen SYBR Green PCR kit and a Biorad DNA Engine Opticon 2 system. The threshold cycle (CT) was determined as the cycle with a signal higher than that of the background plus 10 s.d. Catharanthus roseus 40S ribosomal protein S9 (Rps9), encoded by a house-keeping gene, was used to adjust the amount of the totalmRNAin all samples. Real-time RT–PCR was performed in triplicate and the data are pictured as the relative expression levels of rebH and rebF mRNA in transgenic hairy roots as well as hairy roots lacking the pCAMBIA plasmid (Supplementary Fig. 23). |
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找一個(gè)專業(yè)的詞典,如生物醫(yī)藥大辭典,就可以自己解決了啊。www.bioon.cn上有在線的....您完全可以自己搞定的.. |
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評估生物堿的穩(wěn)定性在后續(xù)生產(chǎn)氯化 亞文化群。從十根小費(fèi)毛狀根pCAMRebH / RebF變形 pCAMPyrH / RebF / STRvm,每3周都在Gamborg subcultured的維他命B5 固體介質(zhì)(half-strength基底鹽、維生素、30克l21告誡人們,蔗糖, 每晚l21瓊脂、pH 5.7)。達(dá)到了26 uC在黑暗中。生物堿中分離的 分析了毛狀根的21-day-old如上所述,無花果成都(補(bǔ)充)。 RebH檢定的表達(dá),RebF,STRvm酶和PyrH實(shí)時(shí) 逆轉(zhuǎn)錄聚合酶鏈反應(yīng)技術(shù)。 實(shí)時(shí)逆轉(zhuǎn)錄聚合酶鏈反應(yīng)技術(shù)是用來評估你的表達(dá)水平 RebH和RebF。毛狀根的表達(dá)水平。rhizogenes感染a 缺乏pCAMRebH / RebF構(gòu)建進(jìn)行了比較以表達(dá)水平 在pCAMRebH / RebF毛狀根的與眾不同。信使RNA從轉(zhuǎn)變 毛狀根的是分離和純化DNA使用Qiagen從污染物 RNeasy Rnase-free DnaseI迷你工具及植物,分別。結(jié)果 當(dāng)時(shí),一mRNA reverse-transcribed Qiagen QuantiTect使用 逆轉(zhuǎn)錄組件,然后受到PCR與特定的底漆(見 SYBR Qiagen下面),是一種綠色PCR球衣,同時(shí)有Biorad DNA引擎Opticon 2 系統(tǒng)。周期(CT)的閾值,確定了其信號(hào)的周期下更高 比加10 s.d的背景。Catharanthus roseus學(xué)生9(Rps9 40多歲的(像是核糖體蛋白質(zhì)基因),編碼,是由一個(gè)做家務(wù)的問題基因用于調(diào)整金額... 僅供參考 |
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