Construction method of double arginine pathway protein secretion vector and use thereof

Abstract

The invention relates to a building method of a double-arginine approach protein secretion vector and applications thereof. In the invention, a Tat approach signal peptide from a Bacillus subtilis is adopted for building the vectors of pMKSig and pMASig of a folded protein or a non-secretory protein in a secretory expression cell, moreover, a heat-resistance-galactosidase from a Bacillus stearothermophilus is used as a target protein example for attempting the workability of the built secretion vector. The contrast detection on the activity of the enzymes inside and outside the cell is carried out on the secretion of the heat-resistance-galactosidasein in the Bacillus subtilis realized by the two vectors,and the protein band expressed by secretion is displayed by a denatured polyacrylamide protein electrophoresis. The invention effectively solves the secretion of the folded (incapable of secretion) protein in the cell in the Bacillus subtilis. The technology can be popularized in the secretion expression of other proteins which are incapable of secretion, thereby having excellent application prospects.

Description

A construction method and application of a double arginine pathway protein secretion vector 【Technical field】 The invention relates to the technical fields of microbial molecular biology and genetic engineering, and relates to the secretion and expression of microbial proteins. 【Background technique】 After organisms synthesize certain proteins, they can export these proteins outside the body through specific mechanisms, and this export process is called secretion. For industrial-level recombinant expression aimed at increasing the yield of exogenous proteins, the secretion method is often used to export the target protein outside the cell, which can greatly simplify the downstream protein separation and purification process and save operating costs. Therefore, protein secretion has always been an important aspect of biotechnology research. A hotspot of (TjalsmaH, BolhuisA, JongbloedJD, et al. Signal peptide-dependent protein transport in Bacillus subtilis: agenome-based su...

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This increases the production cost of these enzymes to some extent

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