Space enterococcus faecium LCT (Liquid-based Cytology Test )-EF301
A technology of LCT-EF301, Enterococcus faecium, applied in the field of microorganisms, can solve the problems that have not been reported, limitations and other issues.
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Embodiment 1
[0018] Embodiment 1, hemolysis test of Enterococcus faecium LCT-E F301
[0019] The hemolysis test adopts spotting method. The strains are spot-planted on the blood agar medium, kept at a constant temperature of 37°C, and cultured upside down. As a result, there is no hemolysis pattern around the colony of Enterococcus faecium LCT-E F301, see figure 1 . It can be seen from the figure that Enterococcus faecium LCT-EF301 has no hemolysis phenomenon.
Embodiment 2
[0020] Embodiment 2, the biolog test of Enterococcus faecium LCT-EF301
[0021] 1) Bacterial suspension preparation: Take out the slant surfaces of the space strain Enterococcus faecium LCT-E F301 and the ground control strain, dip the bacteria with a sterilized bamboo stick and inoculate them in LB liquid medium (4ml system) for 24 hours. Then take 2ml of bacterial liquid at 6000r / s, centrifuge for 5min, discard the supernatant medium, and leave the centrifuged bacterial cells. Wash the cells with sterilized normal saline, centrifuge at 6000r / s for 5min, discard the supernatant again, suspend the cells with Biolog inoculum, and finally transfer to 15ml inoculum to adjust the concentration to about 107-108.
[0022] 2) Inoculation: Pour the above bacterial suspension into the inoculation dish, oscillate evenly, add the sample with a 12-channel (or 8-channel) row gun, add 100ul to each Biolog plate hole, a total of 96 wells, after adding the sample, cover Cover the Biolog plat...
Embodiment 3
[0025] Example 3. Genome, transcriptome sequencing and proteomic analysis of Enterococcus faecium LCT-EF301
[0026] The whole genome sequence of Enterococcus faecium LCT-EF301 was performed using high-throughput Illumina sequencing technology. The whole genome sequence of Enterococcus faecium LCT-EF301 includes 14 Scaffolds. The whole genome sequence is 2,696,027bp long and has 1 SNP site. type, with a GC content of 37.98%, and 2,639 potential protein-coding genes.
[0027] After extracting the total RNA of Enterococcus faecium LCT-EF301 strain, use the kit to remove the rRNA, add fragmentation buffer to break the mRNA into short fragments, and use the mRNA as a template to synthesize the first cDNA strand with six base random primers (random hexamers) , then add buffer, dNTPs, RNase H and DNA polymerase l to synthesize the second cDNA strand, after purification by QiaQuick PCR kit and elution with EB buffer, do end repair, add polyA and connect the sequencing adapter, and th...
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