提高蛋白质表达的各种启动子的筛选

IF 0.6 Q4 FOOD SCIENCE & TECHNOLOGY

Journal of microbiology, biotechnology and food sciences Pub Date : 2023-07-31 DOI:10.55251/jmbfs.9863

Nandini B NATARAJ, R. Sudhakaran

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引用次数: 0

摘要

启动子是在大肠杆菌中表达重组蛋白的基本元件之一。它是驱动重组蛋白生产表达的DNA序列。随着启动子的选择，蛋白质参数、诱导温度、诱导剂、诱导条件和蛋白质溶解度等因素对靶蛋白的产量起着重要作用。在本研究中，我们评估了三种不同启动子（phoA、cstA和pLpR）的强度，并将其与已知的强启动子之一T7进行了比较。构建了一个含有这三个启动子的载体，用于表达FCTRX 1–15标记的胰岛素原基因。用色谱法分离、溶解和纯化表达的蛋白质。比较了从不同启动子驱动的构建体获得的胰岛素原的产量，并且观察到从温度诱导型pLpR系统获得的包涵体（IB）产量与T7系统相当，并且这可以通过优化发酵条件来进一步提高。

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SCREENING OF VARIOUS PROMOTERS FOR INCREASED PROTEIN EXPRESSION

One of the essential elements in the in expression of recombinant proteins in Eshecichia coli is a promoter. It is a sequence of DNA that drives the expression of recombinant protein production. Along with the choice of promoter a number of factors such as protein parameters, induction temperature, inducer, induction conditions and protein solubility play a major role in deciding the yield of target protein. In the present study, we have evaluated the strength of three different promoters (phoA, cstA and pLpR) and compared it with one of the strong promoters known, T7. A vector with these three promoters were constructed for expression of FCTRX 1 – 15 tagged proinsulin gene. Protein expressed was isolated, solubilized and purified using chromatography. Yields of proinsulin obtained from different promoter driven constructs was compared and it was observed that the inclusion bodies (IB) yield obtained from temperature inducible pLpR system was comparable with T7 system and this can be further increased by optimizing the fermentation conditions.

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来源期刊

Journal of microbiology, biotechnology and food sciences FOOD SCIENCE & TECHNOLOGY-

CiteScore

1.90

自引率

0.00%

发文量

156

审稿时长

8 weeks

期刊介绍： The Journal of Microbiology, Biotechnology and Food Sciences is an Open Access, peer-reviewed online scientific journal published by the Faculty of Biotechnology and Food Sciences (Slovak University of Agriculture in Nitra). The major focus of the journal is regular publishing of original scientific articles, short communications and reviews about animal, plant and environmental microbiology (including bacteria, fungi, yeasts, algae, protozoa and viruses), microbial, animal and plant biotechnology and physiology, microbial, plant and animal genetics, molecular biology, agriculture and food chemistry and biochemistry, food control, evaluation and processing in food science and environmental sciences.