Plasmid vector of escherichia coli secretory expression heterologous protein and establishment method of plasmid vector

Abstract

The invention discloses a plasmid vector of escherichia coli secretory expression heterologous protein and an establishment method of the plasmid vector, and in particular discloses a recombinant secreting type expression vector. The vector comprises an alkaline phosphatase PhoA secretory signal peptide and multi-poly histidine sequence, and can express the heterologous recombinant protein through IPTG (Psopropyl-beta-d-Thiogalactoside) induction in escherichia coli BL21. The invention further discloses techniques for establishment of the vector, PhoA induced peptide-heterologous protein fusion gene clone and fusion protein expression and detection. As the heterologous protein is enabled to be stably expressed in an extracellular manner, the vector can be applied to secretory expression of ordinary protein.

Description

technical field [0001] The invention relates to the technical field of protein expression and purification, in particular to a plasmid vector for secreting and expressing foreign proteins and a construction method thereof. technical background [0002] Protein expression and purification technology has important applications in many fields such as low-cost extraction of enzymes and active proteins, protein function research, and protein structure analysis. The most basic requirement for the purified protein is that the protein can be well dispersed in the corresponding solution and have good biological activity. At present, most of the conventional expression vectors are based on the intracellular expression technology of E. coli, which has the advantage that E. coli is easy to cultivate and disrupt, which is beneficial to the purification of proteins. The disadvantage is that the expressed exogenous protein is limited to the inside of the cell, and the protein yield of the...

AI Technical Summary

Problems solved by technology

The disadvantage is that the expressed foreign protein is limited to the inside of the cell. Due to the limited space, the protein yield of the soluble part is limited, and many proteins form aggregation or inclusion bodies in the cell, which requires further denaturation and renaturation treatment.

However, proteins in the form of aggregation or inclusion bodies are often difficult to redisperse in a suitable system due to various molecular modifications.

When looking for renaturation conditions, more conditions are required to explore the process, resulting in cost consumption

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