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After cut down by restriction enzyme,the dhaT gene with own ribosome binding site was inserted into recombining plasmid pMD19-T Simple/gldABC in tandem. Thereafter,the gene of gldABC and dhaT was subcloned into expression vector pET28a(+).
中文意思:
将限制性内切酶处理后的含有自身核糖体结合位点的dhaT基因插入到质粒pMD19-T Simple/gldABC中gldABC基因的下游,形成重组克隆质粒pMD19-T Simple/gldABC-dhaT,并进一步将gldABC与dhaT串联基因亚克隆到表达载体pET28a(+)上。


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After 10 h cultivation at 30°C and 5 h induction cultivation at 42°C, the final cell density reached 60 OD600, rhTNFaDK2 was more than 50% of the total amount of protein in E. coli, near 2 g pure rhTNFαDK2 per liter broth was obtained. 在5L在NBS-Bioflo3000型自动控制发酵罐中采取加速补料的补料分批培养,重组大肠杆菌YK537/pSB-TK经10h30°C培养和5h42°C诱导培养,最终密度达到60OD600,rhTNFα-DK2的表达量占菌体总蛋白的50%以上,每升发酵液纯化可得到近2g的rhTNFα-DK2。
After 20 hours, H_2O_2 was added to final concentration 1%, then culture to 24 hours. In this condition, the biomass was 5.26g/100ml, SOD activity was 2662u/g fresh cell, SOD production was 1.40×10~4u/100ml, which was 99% high than original strain.在此条件下,细胞生物量为5.26g/100ml,SOD含量为2662u/g湿菌体,SOD产量为14002u/100ml,产量较出发菌提高99%。
After 3 days 2.5μg/mL of arabinoside cytarabine(Ara-C) was added to the culture for 24h to inhibit the outgrowth of glial cells,mechanocytes and nerve stem cells,etc.应用2.5μg/mL阿糖胞苷(Ara-C)对培养3 d的神经元进行24 h干预,去除神经胶质细胞、成纤维细胞、神经干细胞等杂质细胞,以提高神经元产率;
After cut down by restriction enzyme,the dhaT gene with own ribosome binding site was inserted into recombining plasmid pMD19-T Simple/gldABC in tandem. Thereafter,the gene of gldABC and dhaT was subcloned into expression vector pET28a(+).将限制性内切酶处理后的含有自身核糖体结合位点的dhaT基因插入到质粒pMD19-T Simple/gldABC中gldABC基因的下游,形成重组克隆质粒pMD19-T Simple/gldABC-dhaT,并进一步将gldABC与dhaT串联基因亚克隆到表达载体pET28a(+)上。
After diapause the RNA/DNA ratio and the protein content arise to 7.0 and 133 pig/rag respectively.滞育结束后RNA/DNA比值上升为7.04,蛋白含量为133微克/毫克脂肪体。 成虫繁殖期与滞育期不同阶段脂肪体核酸、蛋白量的差异反映卵黄蛋白合成水平。
After injecting mifepristone, Thl-type cytokines were high remarkably comparing with the control experiment while the Th2-type cytokines changing little.米非司酮促流产后,Th1型细胞因子的表达明显升高,与对照组相比差异极显著,Th2型细胞因子含量仅在用药12h后有显著下降,但很快升高到正常水平;
After the Symphyso-collection (278 genes) of the 300 genes wasgot, cluster analysis was done by Cluster software and TreeView.对这 300 个基因取并集(共 278 个基因),用 Cluster 软件及 TreeView 完成系统聚类分析。
After the cultivation for 26h at 40℃; in a 100L fermentor, a spore concentration of 1.07×1010 CFU/mL was obtained with a spore percentage of greater than 93% and a maximum live cell concentration of 1.14×1010CFU/mL.当在100L发酵罐上40℃培养26h时,得到的菌体浓度为1.14×10~(10)CFU/mL,芽孢浓度为1.07×10~(10)CFU/mL,芽孢形成率为93%。
After the floating neurosphere was formed in primary culture, some neural stem cells were used for suspension culture whereas part of cells was cultured adhesively to the flask precovered by 0.001% polyornithine and 0.2% gelatin.原代培养出现大量悬浮神经干细胞球后,一部分细胞继续悬浮培养,另一部分细胞种入用0.001%多聚鸟氨酸和0.2%明胶包被的培养瓶中贴壁培养。
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