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求翻译一段文字 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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老男人(金币+5, 翻译EPI+1): 确实只能仅供参考。。。 2011-03-27 13:39:45
ringzhu(金币-5, 翻译EPI-1): 收回EPI 本版严禁机器翻译~~ 2011-03-27 18:06:56
老男人(金币+5, 翻译EPI+1): 确实只能仅供参考。。。 2011-03-27 13:39:45
ringzhu(金币-5, 翻译EPI-1): 收回EPI 本版严禁机器翻译~~ 2011-03-27 18:06:56
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评估生物碱的稳定性在后续生产氯化 亚文化群。从十根小费毛状根pCAMRebH / RebF变形 pCAMPyrH / RebF / STRvm,每3周都在Gamborg subcultured的维他命B5 固体介质(half-strength基底盐、维生素、30克l21告诫人们,蔗糖, 每晚l21琼脂、pH 5.7)。达到了26 uC在黑暗中。生物碱中分离的 分析了毛状根的21-day-old如上所述,无花果成都(补充)。 RebH检定的表达,RebF,STRvm酶和PyrH实时 逆转录聚合酶链反应技术。 实时逆转录聚合酶链反应技术是用来评估你的表达水平 RebH和RebF。毛状根的表达水平。rhizogenes感染a 缺乏pCAMRebH / RebF构建进行了比较以表达水平 在pCAMRebH / RebF毛状根的与众不同。信使RNA从转变 毛状根的是分离和纯化DNA使用Qiagen从污染物 RNeasy Rnase-free DnaseI迷你工具及植物,分别。结果 当时,一mRNA reverse-transcribed Qiagen QuantiTect使用 逆转录组件,然后受到PCR与特定的底漆(见 SYBR Qiagen下面),是一种绿色PCR球衣,同时有Biorad DNA引擎Opticon 2 系统。周期(CT)的阈值,确定了其信号的周期下更高 比加10 s.d的背景。Catharanthus roseus学生9(Rps9 40多岁的(像是核糖体蛋白质基因),编码,是由一个做家务的问题基因用于调整金额... 仅供参考 |
3楼2011-03-23 21:24:26
2楼2011-03-23 20:52:27
