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serratia marcescens LCT-SM166 strain in space environment

A technology of LCT-SM166, space environment, applied in the field of protein molecules

Inactive Publication Date: 2013-07-03
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies on Serratia discoloration in space environment have not been reported yet

Method used

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  • serratia marcescens LCT-SM166 strain in space environment
  • serratia marcescens LCT-SM166 strain in space environment
  • serratia marcescens LCT-SM166 strain in space environment

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0022] Specific implementation example 1, analysis of phenotypic characteristics of LCT-SM262:

[0023] (1). Morphological characteristics: take 50ul of the sample and centrifuge, discard the supernatant, add 50ul of sterile water, absorb 20ul and fix it on the glass slide; Carry out the initial dyeing on the sample for 1min, then rinse the dye solution with tap water; for mordant dyeing, absorb 2 drops of Gram stain II (iodine solution) to cover the painted surface and dye for about 1min, then rinse the dye solution with tap water, and absorb the dye on the coated surface with absorbent paper. Moisture; for decolorization, absorb 2 drops of Gram stain III (alcohol) on the painted surface for about 30 seconds, rinse the glass slide with water, and absorb the water on the painted surface with absorbent paper; for counterstaining, absorb Gram stain IV ( Red) 2 drops on the coated surface for about 1min, rinse the glass slide with water, and absorb the water on the coated surface...

specific Embodiment 2

[0035] Specific implementation example two, the genome and comparative genomics of LCT-SM166:

[0036] Cultivate Serratia discoloration and extract genomic DNA. First, use ultrasonic method to randomly interrupt the qualified DNA samples to obtain a series of DNA fragments. After treatment with T4 DNA polymerase, Klenow DNA polymerase and T4 polynucleotide kinase, etc. , recover the target fragment, obtain the sequencing library, and use Illumina’s Genome Analyzer II for sequencing; the reads obtained after sequencing are assembled and analyzed after filtering; use G|immer3.0 software to annotate genes and analyze SNP, Indel, and gene weight row and structural variation, etc. For the gene sequencing quality control of LCT-SM262, see Figure 4 , Figure 5 . The SNP altered genes are shown in Table 2, and Indel analysis did not reveal significant changes. In addition, the LCT-SM166 prodigiosin synthesis gene (pigB-pigP) in the space environment, the metabolite prodigiosin en...

specific Embodiment 3

[0039] Specific implementation example three, transcriptome sequencing analysis of LCT-SM262 strain:

[0040] After extracting the total RNA of the LCT-SM262 strain, the kit was used to remove the rRNA. Add fragmentation buffer to break mRNA into short fragments, use mRNA as a template, use six base random primers (random hexamers) to synthesize the first cDNA strand, then add buffer, dNTPs, RNase H and DNA polymerase I to synthesize the second strand The cDNA chain is purified by the QiaQuick PCR kit and eluted with EB buffer, then repaired at the end, added polyA and connected to the sequencing adapter, then used agarose gel electrophoresis to select the fragment size, and finally performed PCR amplification. The sequencing library was sequenced using Illumina HiSeq2000. After removing the impurity data from the original sequence data, count rRNA statistics, evaluate the distribution of Reads on the genome, gene coverage, count gene expression differences, analyze different...

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Abstract

The invention relates to a serratia marcescens LCT-SM166 strain in a space environment, as well as the biological characteristics, the genomic DNA sequence, the transcriptome and the differential proteomics thereof, particularly to the functional sequences and the molecules acquired through analysis and identification compared with serratia marcescens on the ground. Therefore, the function and mechanism research of microorganisms in the space environment is facilitated, and the space environmental safety is guaranteed; discoveries of novel virulence and drug-resistant target spots are facilitated, and novel treatment target spots can be provided for clinical infectious diseases; and prodigiosin as the metabolite has the anti-tumor and immunosuppressive functions, thereby having a potential application prospect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a new microbial strain and its biological characteristics, genome DNA sequence, transcriptome and differential proteomics, using bioinformatics technology to obtain its comparative genomics information, transcriptome sequencing and analysis of differences Expressed genes and protein molecules identified by differential proteomics. Background technique [0002] With the implementation of my country's huge space programs such as the Shenzhou spacecraft and the space station, the demand for medical support for manned spaceflight has become increasingly prominent. During spaceflight activities, some microorganisms can enter space with the human body or aircraft. Various bacteria including Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Serratia, Bacillus cereus and Enterococcus have been detected on the space station. The natural space environment ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/425
Inventor 刘长庭常德王雅娟方向群李天志王俊峰郭英华苏龙翔陈振鸿刘岩
Owner GENERAL HOSPITAL OF PLA
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