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首页> 外文期刊>Annals of microbiology >The role of lac operon and lac repressor in the induction using lactose for the expression of periplasmic human interferon-α2b by Escherichia coli
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The role of lac operon and lac repressor in the induction using lactose for the expression of periplasmic human interferon-α2b by Escherichia coli

机译:lac操纵子和lac阻遏物在乳糖诱导大肠杆菌表达周质人干扰素-α2b中的作用

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The effect of lac operon in the induction using lactose for the expression of periplasmic human interferon-α2b (PrIFN-α2b) was studied in shake flask culture. Escherichia coli strains Rosetta2 (DE3) [R2 (DE3)] containing the lac operon and Rosetta-gami2 (DE3) [RG2 (DE3)] containing the deletion of entire lac operon with high level of lac repressor were used. R2 (DE3) over-expressed PrIFN-α2b at substantial levels (270–380?μg/L) in lactose-induced media. In spite of the deletion of lac operon in RG2 (DE3), the cells exposed to lactose produced PrIFN-α2b albeit in less quantity (18–20?μg/L). Under similar conditions, the percentage of IFN-α2b translocated into periplasm for cells induced with lactose was 43–57 and 15–30% in R2 (DE3) and RG2 (DE3), respectively. The PrIFN-α2b expressed by RG2 (DE3) grown in control medium and Terrific broth was 290.3 and 134.7?μg/L, respectively. The basal expression levels obtained in R2 (DE3) strain were 10-fold higher than those obtained in RG2 (DE3) strain. The target proteins expressed in uninduced state did not affect the growth, indicating that IFN-α2b was non-toxic to the bacterial cells. Equivalent level of PrIFN-α2b expression was obtained in RG2 (DE3) induced by IPTG and in R2 (DE3) induced by lactose.
机译:在摇瓶培养中研究了lac操纵子在乳糖诱导的周质人干扰素-α2b(PrIFN-α2b)表达中的作用。使用含有lac操纵子的大肠杆菌菌株Rosetta2(DE3)[R2(DE3)]和含有高水平的lac阻抑剂的缺失整个lac操纵子的Rosetta-gami2(DE3)[RG2(DE3)]。 R2(DE3)在乳糖诱导的培养基中大量表达(270–380?μg/ L)的PrIFN-α2b。尽管RG2(DE3)中的lac操纵子缺失,但暴露于乳糖的细胞产生的PrIFN-α2b数量较少(18–20?g / L)。在类似条件下,乳糖诱导的细胞中转移到周质中的IFN-α2b的百分比在R2(DE3)和RG2(DE3)中分别为43–57和15–30%。 RG2(DE3)在对照培养基和Terrific肉汤中生长的PrIFN-α2b分别为290.3和134.7μg/ L。 R2(DE3)菌株获得的基础表达水平比RG2(DE3)菌株获得的基础表达水平高10倍。以未诱导状态表达的靶蛋白不影响其生长,表明IFN-α2b对细菌细胞无毒。在由IPTG诱导的RG2(DE3)和由乳糖诱导的R2(DE3)中获得了等价的PrIFN-α2b表达。

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