Escherichia coli-bacillus subtilis shuttle expression vector and application thereof

Abstract

The invention discloses an Escherichia coli-bacillus subtilis shuttle expression vector and application thereof, and belongs to the field of genetic engineering. The Escherichia coli-bacillus subtilis shuttle expression vector comprises a bacillus subtilis promoter sequence, a bacillus subtilis signal peptide coding sequence, an artificially designed ribosome binding sequence which can be identified by bacillus subtilis, the artificially designed enzyme cutting site of bacillus subtilis signal peptidase, an artificially designed multiple cloning site, a bacteriophage terminator sequence, a bacillus subtilis replicating initial sequence, and an Escherichia coli replicating initial sequence. The vector can be replicated in Escherichia coli and can also be replicated and expressed in the bacillus subtilis, so that vectors for a bacillus subtilis expression system are enriched; and the vector uses a B.subtilis deacetylated chitinase signal peptide for the first time, the signal peptide has high capability of guiding to secrete extracellular protein, a reporter gene is aspergillus nidulans pectate lyase gene, and the extracellular expression level reaches 600U / mL which is higher than that when the pectate lyase gene uses other vectors.

Description

technical field

[0001] The invention designs an Escherichia coli-Bacillus subtilis shuttle expression vector and its application, belonging to the field of genetic engineering. Background technique

[0002] In the research on the expression of enzymes used in the food industry and industrial fermentation production, the recombinant expression of food enzymes with microbial expression systems has broad application prospects. At present, the E. coli expression system and yeast expression system have convenient sources. Many research institutions have developed a variety of expression vectors and expression host bacteria over the years, and have modified the E. coli expression system and yeast expression system in multiple directions. However, many problems in the E. coli expression system still exist, such as the formation of inclusion bodies and the toxicity of E. coli expression host bacteria. Yeast expression system The expression system has unique advantages because it ca...

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