Construction method of recombinant plasmid pNZ8048-plnC

Abstract

The invention discloses a construction method of a recombinant plasmid pNZ8048-plnC. A Lactobacillus plantarum ZJQ genome is used as a template, a PCR amplification is carried out for a plnC target gene, specifically amplified strips of amplified gene fragments are purified by a PCR product purifying kit, a double digestion is carried out for the purified PCR products and a plasmid pNZ8048 with NcoI and Hind III, after purifying, an overnight connection is carried out at 16 DEG C, and the recombinant plasmid pNZ8048-plnC is obtained. The constructed recombinant plasmid pNZ8048-plnC can be expressed in Lactobacillus plantarum ZJ316 (lack of regulatory protein PlnC and PlnD). The system is helpful for researching and discussing a transcription regulation mechanism of PlnC in the Lactobacillus plantarum ZJ316, and furthermore is helpful for revealing an anabolism regulation mechanism of PlnC to Lactobacillus plantarum ZJ316 bacteriocin; the system provides a theoretical basis for a Lactobacillus plantarum ZJ316 quorum-sensing mode, and provides an innovative thinking for high-yield research of Lactobacillus plantarum ZJ316 bacteriocin; the system has important meanings for development and business utilization of Lactobacillus plantarum ZJ316 bacteriocin.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering, in particular to a construction method of a recombinant plasmid pNZ8048-plnC. Background technique [0002] Lactobacillus plantarum is widely present in human and animal intestines, preserved fruits and vegetables, and preserved meat products, and can utilize glucose fermentation and metabolism to produce a large amount of lactic acid. In addition, Lactobacillus plantarum can also secrete some bacteriocin with antibacterial activity extracellularly. Bacteriocins can be adsorbed on the cell membrane of the indicator bacteria, causing the cell membrane structure of the indicator bacteria to change to form voids, resulting in the leakage of K+, ATP, amino acids and phosphates in the indicator bacteria cells and death. Bacteriocin has the characteristics of thermal stability, non-toxicity, high efficiency and safety. [0003] Lactobacillus plantarum ZJ316 is a strain of Lactobacillus ...

AI Technical Summary

Problems solved by technology

Through the analysis of the quorum-sensing regulatory operon plnABCD of Lactobacillus plantarum bacteriocin biosynthesis, Lactobacillus plantarum ZJ316 has the signal peptide gene gene plnc8IF and the histidine protein kinase gene plnB regulated by quorum sensing, but lacks the regulatory protein gene in the cell plnC and plnD, therefor

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