Yeast extract mediated autoinduction of lacUV5 promoter: an insight
Yeast extract mediated autoinduction of lacUV5 promoter: an insight
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DOI：
10.1016/j.nbt.2009.08.002
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发表时间：
2009-12-31
影响因子：
5.4
通讯作者：
Padmanabhan, Sriram
中科院分区：
文献类型：
--
作者：
Nair, Rahul;Salvi, Pankaj;Padmanabhan, Sriram
我们报告了一种简单而经济的自诱导培养基成分，负责在大肠杆菌lacUV 5启动子系统下自诱导蛋白质。高浓度的酵母提取物（YE）能刺激BL 21（DE 3）细胞T7 RNA聚合酶的表达，而对BL 21 A1细胞无此作用。对不同浓度的YE的影响的系统研究表明，人粒细胞集落刺激因子（rhGCSF）、人干扰素α 2b（rhIFN-α 2b）和葡激酶（rSAK）在不存在任何特异性诱导剂如IPTG或另外的乳糖的情况下在BL 21（DE 3）细胞中的表达。对YE的诱导组分的进一步研究揭示了存在大量内源性乳糖作为所观察到的自诱导现象的贡献因素。本文强调了在生物技术工业中使用本媒体组件进行大规模生产的易扩展性。
We report a simple and cost-effective autoinducible media component responsible for the autoinduction of proteins in Escherichia coli under lacUV5 promoter system. Yeast extract (YE) at high concentration was found to stimulate the expression of T7 RNA polymerase in BL21(DE3) cells while such an effect was not seen in BL21A1 cells. A systematic study on the effect of varying concentrations of YE indicated several folds higher expression of genes viz., human granulocyte colony stimulating factor (rhGCSF), human interferon alpha 2b (rhIFN-alpha 2b) and Staphylokinase (rSAK) in BL21(DE3) cells in the absence of any specific inducer like IPTG or additional lactose. Additional investigations on the inducible component of the YE revealed the presence of significant amount of endogenous lactose as the contributory factor for the observed autoinduction phenomenon. This paper highlights the easy scalability of the use of the present media component for large-scale production in biotechnology industry.