Broad-spectrum anti-phage E. Coli BL21-(DE3)-PR and application thereof
A technology of Escherichia coli, bacteriophage, applied in the field of bioengineering
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Embodiment 1
[0046] Example 1. Screening and preservation of anti-phage strain E.coli BL21(DE3)-PR
[0047] 1. Screening of anti-phage strain E.coli BL21(DE3)-PR
[0048] 1. Preparation of BL21(DE3) bacterial solution
[0049] E.coli BL21(DE3) was inoculated in LB liquid medium and cultured overnight at 37°C and 220rpm to obtain BL21(DE3) bacterial liquid.
[0050] 2. Isolation of phage
[0051] Take sewage (the sewage comes from the waste water of the 307 Hospital of the Chinese People's Liberation Army), centrifuge at 10000r / min for 10min, and collect the supernatant. The supernatant was filtered with a 0.22 μm filter to obtain a sewage filtrate. Then add 200 μL of sewage filtrate and 4 mL of BL21 (DE3) bacterial solution prepared in step 1 into 2 mL of 3×LB liquid medium, and culture at 37° C. and 220 rpm for 6 hours to obtain phage stock solution. Then the phage stock solution was filtered with a 0.22 μm microporous membrane to obtain a phage preservation solution, which was stored...
Embodiment 2
[0071] Embodiment 2, performance detection of anti-phage bacterial strain BL21 (DE3)-PR
[0072] 1. Phage plaque test against phage strain BL21(DE3)-PR
[0073] In order to detect the anti-phage performance of the anti-phage strain Escherichia coli BL21(DE3)-PR (hereinafter abbreviated as E.coli BL21(DE3)-PR), a phage plaque test was carried out. The specific steps are as follows: Take 300 μL of the BL21 bacterial solution prepared in Step 1 of Example 1 and 300 μL of the phage-resistant bacterial strain BL21(DE3)-PR screened in Step 1 of Example 1 and add 5 mL of LB semi-solid medium respectively , mix evenly, spread on the LB solid medium plate containing the lower layer, and dry at room temperature for 5 minutes;8 pfu / mL phage preservation solution (T4, vB_EcoM_IME281, vB_EcoM_IME253, vB_EcoM_IME339, vB_EcoM_IME340, vB_EcoM_IME341, T1, vB_EcoS_IME18, vB_EcoS_IME253, vB_EcoM_IME347, vB_EcoM_IME341, T1, vB_EcoS_IME18, vB_EcoS_IME253, vB_EcoS_IME347, JMPW1 bacteria-free cultur...
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