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Controlling method for expression system of T7 induced by laevorotary arabinose

A control method and technology of expression system, which are applied in the control field of L-arabinose-induced T7 expression system, can solve problems such as excessive protein, and achieve the effects of maintaining stability, having economic competitiveness and reducing cost.

Inactive Publication Date: 2004-06-16
WIDETEX BIOTECH
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Problems solved by technology

However, this known technique is to clone the T7 gene 1 regulated by the araBAD promoter on the plastid with the pACYC184 replication origin (origin), so it leads to excessively high protein production when not induced, and is not suitable for producing Potentially Toxic Recombinant Proteins

Method used

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  • Controlling method for expression system of T7 induced by laevorotary arabinose
  • Controlling method for expression system of T7 induced by laevorotary arabinose
  • Controlling method for expression system of T7 induced by laevorotary arabinose

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Embodiment Construction

[0027] The present invention uses L-arabinose to induce the control method of the T7 expression system, comprising the following steps:

[0028] Construct a strain of recombinant strain, this recombinant strain contains a plastid containing T7 promoter, and the chromosome of this recombinant strain contains T7 gene 1 and araC control gene controlled by araBAD promoter;

[0029] The recombinant bacterial species is cultivated by a feed batch fermentation method. In the batch fermentation stage, the provided fermentation liquid contains glucose as a carbon source, and in the feed stage, the provided feed liquid contains glycerol and glucose as a carbon source. enabling high cell density fermentation; and

[0030] After the cells reach a certain high density, an inducer containing L-arabinose is added to make the recombinant strain produce a recombinant protein.

[0031] The control method of the present invention further comprises: the recombinant strain BL21 (BAD) constructed ...

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Abstract

The invention discloses a controlling method for constructing a recombination bacterial BL21(BAD) for evoked preparation of T7 RNA polymerase by levorotation arabinose, the chromosome of the bacterial includes a T7 gene 1 controlled by an araBAD promoter, the T7 RNA polymerase produced by the recombination bacterial can evoke and activate the T7 promotor, thus manufacturing the destination gene product cloned at the downstream of the promotor.

Description

technical field [0001] The present invention relates to a method for controlling the gene expression vector (expression vector) in E. Source and segmented feed batch fermentation strategies for mass production of recombinant proteins. Background technique [0002] The production of recombinant proteins with commercial value or medical use by biological cells is an extremely important and economically forward-looking biotechnology industry. Generally speaking, biological cells that can be used for protein production include microorganisms, insects, animal and plant cells, etc. Among them, the method of using microbial cells to produce recombinant proteins is more economically competitive because microbial cells are easier to produce in large quantities. The formulation of the nutrient matrix required for cultivation, rapid growth and cultivation is simple and inexpensive. Over the past few decades, since many scientific studies have focused on Escherichia coli as the core s...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/67C12N15/70
Inventor 赵云鹏姜中人洪文滨林维兴
Owner WIDETEX BIOTECH
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