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Salinimonas profundi 13199 as well as CRISPR-Cas system and application thereof

The technology of a single-cell bacterium, CGMCC1.17396, is applied to Halomonas abyssalum 13199 and its CRISPR-Cas system and application field, which can solve the problems of class2 complexity and obstacles to the development and application of CRISPR-Cas system

Active Publication Date: 2021-01-01
HOHAI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, class2 using pleiotropic proteins is more complicated, which hinders the development and application of this type of CRISPR-Cas system to a certain extent.

Method used

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  • Salinimonas profundi 13199 as well as CRISPR-Cas system and application thereof
  • Salinimonas profundi 13199 as well as CRISPR-Cas system and application thereof
  • Salinimonas profundi 13199 as well as CRISPR-Cas system and application thereof

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Embodiment

[0039] 1. The bacterial strain containing the CRISPR-Cas system of the present invention Salinimonas profundi Separation method of 13199:

[0040] Take 2 g of the sediment sample, place it in a 50 mL Erlenmeyer flask filled with 18 mL of sterile artificial seawater (containing 3.0% sea salt), shake it on a shaker at 28 °C at 180 rpm for 30 min, and obtain the bacterial suspension ( is 10 -1 The soil dilution solution, that is, the sample solution diluted 10 times); Then use 10 times serial dilution method to dilute successively to obtain 10 -3 and 10 -4 Concentration gradient dilution; then from 10 -4 Take 0.1 mL of the concentration gradient suspension and spread it on the PYGV recovery medium supplemented with 2.5% sea salt with a sterile applicator. Each 1000 mL of PYGV medium contains 20 mL of mineral salt solution, 0.25 g of peptone, 0.25 g of yeast extract, 15 g of agar powder, pH 7.0-7 2. Sterilize at 121°C for 20 minutes, and add 2.5% when cooling to about 60°C 1...

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Abstract

The invention discloses salinimonas profundi 13199. The salinimonas profundi has the preservation number being CGMCC1.17396 and a classification name being Salinimonas profundi. The strain is a new bacterial species derived from deep sea, and contains a special CRISPR-Cas system and a large number of heavy metal resistance genes. The sequence homologies of the Cas protein and a protein which is included in a high-quality protein database UniProtKB / Switch-Prot subjected to a certain degree of functional researches are lower than 65%, and possibly have different activities and characteristics. The strain Salinimonas prodi 13199 and the CRISPR-Cas system thereof are novel materials for developing and utilizing the CRISPR-Cas system and performing related researches, and furthermore, the strain has a certain application prospect in the aspects of heavy metal polluted environmental modification preparations.

Description

technical field [0001] The present invention relates to a deep-sea-derived bacterial strain, specifically a strain of Halomonas abyssalus 13199 containing a CRISPR-Cas system and its CRISPR-Cas system and application. Background technique [0002] Clustered regularly interspaced short palindromic repeats and its associated proteins (Clustered regularly interspaced short palindromic repeats-CRISPR associated proteins, CRISPR-Cas) system is a special DNA sequence in the genome of prokaryotes, found in about 84% of archaea and 47 % of bacteria. A complete CRISPR-Cas system contains three functional elements: a series of Cas genes, a leader and a CRISPR array. [0003] Leader is generally located upstream of CRISPR and is an AT-rich region with a length of 300-500 bp, which is considered to be the promoter sequence of CRISPR. The CRISPR array consists of multiple short and highly conserved repeat regions (Direct repeat, DR) and multiple spacers (Spacer). The length of DR is g...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/22C02F3/34B09C1/10C12R1/01C02F101/20
CPCC12N9/22C02F3/34B09C1/10C02F2101/20C12R2001/01C12N1/205Y02W10/10
Inventor 张道锋崔希雯张爱华黄建科薛华朋
Owner HOHAI UNIV
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