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Lactic acid galactococcus vector of food grade

A Lactococcus lactis, food-grade technology, applied in the field of Lactococcus lactis vectors, can solve the problems of restricting the application of Lactococcus lactis gene expression system, inducing signal insecurity, antibiotic genes endangering biological safety, etc.

Inactive Publication Date: 2010-09-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These markers are very important for screening suitable transformants and maintaining the selection pressure of improved genes in genetic engineering, but the transferability of antibiotic genes will endanger biological safety and limit the application of Lactococcus lactis gene expression system in food and pharmaceutical industries
However, the inducible gene expression vector system that has been studied more at present also has inducing signals that are often unsafe environmental stimuli, such as silk mycin C and other unsafe problems. Effectively solving these problems has become a safe and efficient expression drug using microbial genetic engineering. The key to biologically active substances such as vaccines, hormones, growth factors and enzymes

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0027] (1) Bacterial strains and plasmids used and involved in the experiments of the present invention

[0028] 1. Lactococcus lactis LacA (nisin-producing bacteria, the chromosome contains nisI immune gene, isolated from fresh milk according to its immune characteristics to nisin.)

[0029] 2. Lactococcus lactis LacR (nisin-resistant bacteria, the chromosome contains nsr nisin resistance gene, isolated from fresh milk according to its resistance to nisin.)

[0030] 3.Lactococcus lactis Lac6 (Lactococcus lactis with no immunity / resistance to nisin)

[0031] 4. E.coli DH5α (belongs to known technology)

[0032] 5.pSH71 (belongs to known technology)

[0033] (2) The preparation of the medium used and involved in the experiment of the present invention

[0034] 1. LB medium

[0035] Peptone 10.0g

[0036] Yeast powder 5g

[0037] NaCl 10g

[0038] Add distilled water to make up to 1000ml, adjust the pH value to 7.6-7.8 with NaOH, aliquot, autoclave at 121°C for 20min, sto...

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Abstract

The invention discloses a lactic acid galactococcus food vector pN11, with its lactic acid squence being SEQ ID NO:3; it asl odiscloses another lactic acid galactococcus food vector pN12, with its lactic acid squence being SEQ ID NO:4. Said vector in this invention can overcome the antibiotics resistant mark shortcoming of current expression vector.

Description

technical field [0001] The invention relates to a lactococcus lactis carrier, in particular to a food-grade carrier pNI1 and pNI2 of a lactococcus lactis which cannot produce nisin itself and has no nisin resistance. Background technique [0002] Lactic acid bacteria (LAB) is a class of important industrial strains most widely used in food, including more than a dozen genera such as Lactococcus lactis, Lactobacillus, and Bifidobacterium. It is well-known because it has long been used by humans to make kimchi, soy sauce, and to produce cheese and yogurt. It is generally regarded as safe (GRAS) food-grade microorganisms. Lactic acid bacteria have high application value in medicine, industry, agriculture and other important fields closely related to human life. The main functions are: 1) control intestinal infection; 2) increase the nutritional value of certain foods; 3) control serum cholesterol levels ; 4) Improve lactose metabolism; 5) Induce specific and non-specific immu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/74
Inventor 李卫芬周绪霞黄琴
Owner ZHEJIANG UNIV