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Streptococcus agalactiae attenuated strain YM001 genome sequence feature and use thereof

A genome sequence and Streptococcus galactiae technology, applied in the field of Streptococcus agalactiae genome sequence, can solve the problems of indistinguishable, difficult to maintain intellectual property rights and other issues

Inactive Publication Date: 2015-12-16
GUANGXI ACADEMY OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the attenuated strain YM001 obtained by the invention cannot be distinguished from other Streptococcus agalactiae through traditional bacterial identification methods. If the strain is stolen, it is difficult to protect the interests of intellectual property rights

Method used

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  • Streptococcus agalactiae attenuated strain YM001 genome sequence feature and use thereof
  • Streptococcus agalactiae attenuated strain YM001 genome sequence feature and use thereof
  • Streptococcus agalactiae attenuated strain YM001 genome sequence feature and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0010] Example 1 Whole Genome Sequencing and Sequence Characteristics of Streptococcus agalactiae Attenuated Strain YM001

[0011] Genomic DNA of the attenuated Streptococcus agalactiae strain YM001 was extracted, and the library was constructed after passing the test. First, the genomic DNA was randomly interrupted by the ultrasonic method Covaris or Bioruptor to generate a series of DNA fragments with a main band less than or equal to 800bp, and then the sticky ends formed by the interruption were repaired into blunt ends by T4DNAPolymerase, KlenowDNAPolymerase and T4PNK. The base "A" allows the DNA fragment to be connected to a special adapter with a "T" base at the 3' end, and the target fragment ligation product to be recovered is selected by electrophoresis, and then PCR technology is used to amplify the adapter at both ends. DNA fragments; finally, cluster preparation and sequencing with qualified libraries. Basic biological information analysis after disembarkation (1...

Embodiment 2

[0018] Example 2 Characteristics of CRISPRs of Gene Functional Elements of Streptococcus agalactiae Attenuated Strain YM001

[0019] Searching and comparing the CRISPRs of 8 strains of Streptococcus agalactiae (see Table 2) (http: / / crispr.u-psud.fr / Server / ), the results showed that 3 strains of Streptococcus agalactiae serotype Ib were derived from tilapia CRISPRs were not detected, and CRISPR1 was detected in all 5 strains of Ia serotype Streptococcus agalactiae, but CRISPR2 was not detected. The CRISPRs sequences of strains HN016, GD201008-001 and ZQ0910 were consistent and had 8 spacers, while the attenuated strain YM001 had only 7 spacers, and other sequences were consistent with wild strains (see figure 2 ). It shows that it is difficult to eliminate the spacer sequence of CRISPRs of Streptococcus agalactiae under the condition of normal subculture in the wild, but in the absence of the threat of exogenous nucleic acid invasion, the spacer sequence of CRISPRs will be lost...

Embodiment 3

[0022] Example 3 Characteristics of Prophages of Gene Functional Elements of Streptococcus agalactiae Attenuated Strain YM001

[0023] PHAST (http: / / phast.wishartlab.com / index.html) was used to analyze the prophages (Prophages) of HN016, YM001 and GD201008-001, and the results showed that a complete prophage sequence and 1 incomplete prophage sequence, and the coding sequence and position of all prophage sequences are completely identical (see image 3 ), indicating that the prophage sequence of the attenuated Streptococcus agalactiae strain YM001 has not changed, and the complete retention of the prophage sequence is conducive to the integrity of the attenuated strain's antigenicity.

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Abstract

The invention relates to a streptococcus agalactiae attenuated strain YM001 genome sequence feature and a use thereof, especially relates to a streptococcus agalactiae attenuated strain genome sequence feature and concretely relates to streptococcus agalactiae attenuated strain whole genome sequencing, sequence analysis and genome comparison analysis. Compared with a maternal wild virulent strain HN016 genome, the YM001 genome has two large fragment losses, the loss sizes are 5832 bp and 11115 bp, and three rRNA and eleven tRNA gene losses and ten functional gene losses or function damage are caused. Compared with the HN016 genome, the YM001 genome has ten small fragment losses and 28 mononucleotide variants. The YM001 genome variation feature can be used for protection of attenuated strain YM001 intellectual property and construction and screening of a streptococcus agalactiae attenuated strain.

Description

technical field [0001] The present invention relates to the genome sequence of Streptococcus agalactiae (Streptococcus agalactiae), in particular to the characteristics of the genome sequence of the attenuated strain of Streptococcus agalactiae, and in particular to the whole genome sequencing, sequence analysis and genome comparison analysis of the attenuated strain of Streptococcus agalactiae. Background technique [0002] Streptococcus agalactiae, also known as Group B Streptococcus, is a Gram-positive bacterium that, in addition to infecting newborns to cause pneumonia and meningitis, infecting cows to cause mastitis, can also infect reptiles, amphibians and many kinds of fish kind. Streptococcus agalactiae infected tilapia, resulting in the death of tilapia meningoencephalitis, and broke out in the farming areas of China, Brazil, the United States, Thailand and other countries, causing huge economic losses to tilapia farming. The outbreak of Streptococcus agalactiae in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12N1/20C12R1/46
Inventor 王瑞陈明李莉萍甘西梁万文黄婷雷爱莹
Owner GUANGXI ACADEMY OF FISHERY SCI
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