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Vibrio natriegens and application thereof

A technology of sodium vibrio and alginate lyase, applied in the field of microorganisms

Active Publication Date: 2019-09-24
山东海之宝海洋科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Harvard University geneticist George Church and other research groups published related research papers in 2016, pointing out the superiority of natrivibrio compared with Escherichia coli and its feasibility as a new model organism, and developed related genome editing tools, It provides a theoretical basis and technical support for promoting the construction and transformation of the host system of this bacterium, but there is no relevant report in China so far

Method used

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  • Vibrio natriegens and application thereof
  • Vibrio natriegens and application thereof
  • Vibrio natriegens and application thereof

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Effect test

Embodiment 1

[0030] The screening method of embodiment 1 sodium vibrio Vibrio natriegens

[0031] (1) The sea mud was sampled near the kelp breeding factory in Rongcheng City, Shandong Province, and 1g of the sample was taken and dispersed evenly in 50mL of sterile water.

[0032] (2) Inoculate 1 mL of supernatant into 50 mL of screening liquid medium, culture at 28°C and 200 rpm for 2 days, and dilute 10 -6 And spread it on the screening plate medium, culture at 28°C for 2 days, pick single colonies of different shapes and streak the plates several times to obtain pure cultures.

[0033] (3) Pick single colonies of different forms, inoculate them in liquid medium, and culture them at 28°C and 200rpm for 2 days, take the supernatant to measure the enzyme activity of the strain, select a strain with higher enzyme activity, and entrust the China Center for Typical Culture Collection to preserve it. The morphological characteristics, physiological and biochemical characteristics and 16S rDNA...

Embodiment 2

[0034] The identification of embodiment 2 sodium vibrio Vibrio natriegens

[0035] (1) Plate colony morphology

[0036] Such as figure 2 As shown, the plate colony morphology of sodium vibrio SK42.001: Stretch on the plate medium to grow rapidly, and grow a single colony after 24 hours of culture at 28°C. The colony is round and convex, milky white, moist and slightly sticky, and smooth , with flat edges and a diameter of about 0.6-0.8cm.

[0037] (2) The characteristics of the bacteria under the electron microscope

[0038] Such as image 3 As shown, the cell characteristics of the sodium vibrio SK42.001 under the electron microscope: the cell is short, with blunt rounded ends, curved into an arc shape, and the size is 0.6-0.8 μm×1.2-1.4 μm.

[0039] (3) Physiological and biochemical characteristics

[0040] Physiological and biochemical characteristics of sodium vibrio SK42.001: such as figure 1 , Table 1, and Table 2 show that Narophilic Vibrio is Gram-staining negat...

Embodiment 3

[0050] The feasibility of embodiment 3 sodium vibrio Vibrio natriegens as novel model organism

[0051] (1) Measurement of growth rate

[0052] In LB3 liquid medium (NaCl concentration is the LB medium of 3%), it is 16.2min when cultivating generation, while it is 31.4min when referring to bacterial strain E.coli BL21 generation ( Figure 4 ); SK42.001 grew a single colony after 5 hours of streak culture on the LB plate, while E.coli BL21 required 10 hours. It shows that SK42.001 grows rapidly, which is about twice that of E.coli.

[0053] (2) Feasibility of SK42.001 expressing foreign genes

[0054] The broad host plasmid pAWP89 was selected to construct a recombinant plasmid pAWP89-GFP-Cm containing a green fluorescent protein (GFP) reporter gene and a chloramphenicol (Cm) resistance gene selection marker ( Figure 5 ), using the pRK2013 helper plasmid, pAWP89-GFP-Cm was introduced into SK42.001 through triparental conjugation, and the conjugated SK42.001 recombinant bact...

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Abstract

The invention discloses vibrio natriegens and application thereof and belongs to the field of microorganisms. The provided vibrio natriegens SK42.001 grows rapidly, carries natural plasmids, can express exogenous genes, and has the development value and the application potential to serve as a novel model organism. Meanwhile, the strain contains a gene capable of encoding algin lyase and a gene capable of encoding oligo-algin lyase and can also be used for producing and preparing related products of algin oligosaccharides.

Description

technical field [0001] The invention relates to a sodium vibrio and application thereof, belonging to the field of microorganisms. Background technique [0002] Vibrio natriegens is the fastest-growing bacterium known. Under certain conditions, the subculture time is less than 10 minutes, and the growth rate is nearly twice that of the traditional model organism Escherichia coli. Therefore, whether in scientific research or industrial production Among them, the application of sodium vibrio will greatly save time and cost, and it is a new model organism with great potential to replace E. coli. [0003] Harvard University geneticist George Church and other research groups published related research papers in 2016, pointing out the superiority of natrivibrio vibrio compared with Escherichia coli and its feasibility as a new model organism, and developed related genome editing tools, It provides a theoretical basis and technical support for promoting the construction and transf...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/88C12N15/60C12P19/00C12P19/04C12P19/12C12R1/63
CPCC12N9/88C12P19/00C12P19/04C12P19/12C12Y402/02C12N1/205C12R2001/63
Inventor 江波张涛孟青陈静静
Owner 山东海之宝海洋科技有限公司
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