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Fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation and application of fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation

A technology of Fusobacterium nucleatum and strains, applied in the direction of bacteria, gastrointestinal cells, and microbial-based methods, which can solve the problems of rare isolates, unclear composition of Fusobacterium nucleatum subspecies, and difficulty in having sufficient representatives of standard strains sexual issues

Active Publication Date: 2020-03-24
SHANGHAI TENTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The composition of the intestinal flora is diverse, the subspecies composition of Fusobacterium nucleatum in the intestine is not yet clear, it is difficult for a single standard strain to be representative enough, and the isolates of different subspecies isolated from the tumor microenvironment are still rare

Method used

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  • Fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation and application of fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation
  • Fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation and application of fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation
  • Fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation and application of fusobacterium nucleatum subsp.vincentii strain for intestinal tract separation

Examples

Experimental program
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Effect test

Embodiment 1

[0030] This example provides a Fusobacterium nucleatum subsp. vincentii strain THCT14A3 isolated from the intestinal tract. The classification of the strain is named Fusobacterium nucleatum subsp.vincentii, and its preservation number is CCTCC NO: M 2019363, and the preservation date is May 2019 On the 17th, the preservation unit was the China Type Culture Collection Center.

[0031] The 16S rRNA gene sequence of the strain is specifically shown in SEQ ID NO: 1, which has 99.59% identity with the 16S rRNA gene sequence (NCBI accession number NR_113040.1) of F. nucleatum subsp. vincentii strain ATCC 51190, and F. nucleatum subsp. The 16S rRNA gene sequence of .nucleatum strain ATCC25586 (NCBI accession number NR_114702.1) has 98.58% identity.

[0032] The complete genome of THCT14A3 was determined, and it was found that the THCT14A3 genome contained a circular chromosome with a size of 2052554bp and a GC content of 27.46%. It contained 1,898 protein-coding genes and 63 non-codi...

Embodiment 2

[0034] In this example, based on the sequence of THCT14A3, combined with comparative genomics, a specific DNA sequence and corresponding primer sequences that can be used to identify Fusobacterium nucleatum subsp. The specific method is as follows:

[0035] On the basis of the THCT14A3 genome, combined with the Fusobacterium nucleatum genome downloaded from NCBI; after subspecies classification according to the ANIb value; through BLASTn analysis of sequences conserved in the genome of subsp. The genomes were compared with BlASTn, in which they were screened for regions not present in the genomes of other subspecies, ie, subspecies Wenckeii-specific DNA sequences (below). (Because of the inevitable polymorphism in the DNA sequence, it is impossible to exhaustively, the DNA characteristic sequence referred to here includes the following sequences and unlisted fragments with BLASTn similarity to any of the following sequences)

[0036] >vincentii_specific_region

[0037] GGCAA...

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Abstract

The invention provides a fusobacterium nucleatum subsp.vincentii strain THCT14A3 for intestinal tract separation. The classification designation is Fusobacterium nucleatum subsp.vincentii, the preservation number is CCTCC NO:M 2019363, the preservation date is May 17th, 2019, and the preservation unit is China Center for Type Culture Collection. The 16SrRNA gene sequence of the strain is as shownin SEQID NO:1. The invention further provides a specific DNA sequence for identifying the fusobacterium nucleatum subsp.vincentii and a primer sequence corresponding to the specific DNA sequences. Thecolorectal cancer cell line and THCT14A3 co-culture result show that the THCT14A3 can significantly promote colorectal cancer cell proliferation. Therefore, the THCT14A3 provided by the invention canfacilitate micro-ecology research of intestinal tracts suffering from colorectal cancer.

Description

technical field [0001] The invention relates to the field of Fusobacterium nucleatum, in particular to a strain of Fusobacterium nucleatum subsp. Wentenberg isolated from the intestinal tract and its application in the study of intestinal microecology of colorectal cancer. Background technique [0002] Colorectal cancer is not only the third most common cancer in the world, but also the fourth leading cause of cancer death in the world, following lung cancer, liver cancer and gastric cancer. Colorectal carcinogenesis is the malignant proliferation of cells caused by the double blow of genetics and environment. The discovery of the association between gut microbes and colorectal cancer has prompted researchers to investigate the mechanisms of microbial carcinogenesis. At present, a variety of microorganisms associated with colorectal cancer have been discovered, including Citrobacter, Streptococcus bovis, Pasteurella multocida, Helicobacter pylori, human papillomavirus, ente...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N5/09C12Q1/689C12Q1/04C12N15/11C12R1/01
CPCC12N5/0679C12N5/0693C12N2502/70C12Q1/689C12N1/205C12R2001/01
Inventor 秦环龙蔚青毕德玺李豪
Owner SHANGHAI TENTH PEOPLES HOSPITAL
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