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Faded shewanella oneidensis LCT-SM262 strain in space environment

A technology of LCT-SM262, space environment, applied in the field of protein molecules

Inactive Publication Date: 2013-06-26
GENERAL HOSPITAL OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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

Method used

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  • Faded shewanella oneidensis LCT-SM262 strain in space environment
  • Faded shewanella oneidensis LCT-SM262 strain in space environment
  • Faded shewanella oneidensis LCT-SM262 strain in space environment

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment 1

[0027] Specific implementation example 1, analysis of phenotypic characteristics of LCT-SM262:

[0028] 1. Morphological characteristics: Take 50ul sample for centrifugation, discard the supernatant, add 50ul of sterile water, absorb 20ul and fix it on the glass slide; for initial staining, absorb 2 drops of Gram stain I (crystal violet) on the glass slide sample Carry out initial dyeing for 1 minute, 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 1 minute, then rinse the dye solution with tap water, and absorb the water on the coated surface with absorbent paper; For decolorization, absorb 2 drops of Gram stain III (alcohol) on the coated surface for about 30 seconds, rinse the glass slide with water, and absorb the water on the coated surface with absorbent paper; counterstain, absorb Gram stain IV (safranin) Put 2 drops on the coated surface for about 1min, rins...

specific Embodiment 2

[0039] 6. Determination of growth curve: after activation, the strain was inoculated into a 100-hole honeycomb plate, and the growth curve was measured in the Bioscreen, measured for 24 hours, and the data was read. Calculate the average value of the obtained OD value, and then draw the curve, see image 3 . LCT-SM262 showed no difference in growth rate. Specific implementation example two, the genome and comparative genomics of LCT-SM262:

[0040] 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 to sequence; the reads obtained after sequencing are filtered and then assembled and analyzed; Glimmer3.0 software is used to annotate genes and analyze SNPs, Indels, ...

specific Embodiment 3

[0045] Specific implementation example three, transcriptome sequencing analysis of LCT-SM262 strain:

[0046] 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 HiSeq 2000. 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 differen...

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Abstract

The invention relates to a faded shewanella oneidensis LCT-SM262 strain in a space environment, and biological characteristics, a genomic deoxyribonucleic acid (DNA) sequence, a transcriptome and differential proteomics of the faded shewanella oneidensis LCT-SM262 strain, and in particular relates to a functional sequence and a molecule which are analyzed and identified through comparing with ground faded shewanella oneidensis. The mutant gene, differential expression gene and protein molecule of the faded shewanella oneidensis in the space environment can be comprehensively and accurately obtained, so that researches on the function and mechanism of the space environment on microorganisms are facilitated, and the safety of the space environment is guaranteed; novel toxicity and drug-resistant target spots can be conveniently found out, so that novel treatment target spots are provided for clinic infectious diseases; and a metabolic product-prodigiosin of the strain has functions of tumor resisting and immunosuppression and has a potential application prospect.

Description

technical field [0001] The invention belongs to the field of microorganisms and biotechnology, and relates to a new microbial strain and its biological characteristics, genomic DNA sequence, transcriptome and differential proteomics, using bioinformatics technology to obtain its comparative genomics information, transcriptome sequencing analysis Differentially expressed genes and protein molecules identified by differential proteomics. Background technique [0002] With the implementation of huge space programs such as my country's Shenzhou spacecraft and space station, the demand for medical support for manned spaceflight has become increasingly prominent, and the research on space medicine has become increasingly important. Microorganisms are ubiquitous in nature, and space activities are inevitable. Both the human body and space components will carry microorganisms into space. Previous studies on the influence of the space environment on microorganisms have found that in...

Claims

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Application Information

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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徐国纲
Owner GENERAL HOSPITAL OF PLA
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