Electricity transformation method for lactobacillus casei
A technology of Lactobacillus casei and electrotransformation, applied in the field of molecular biology, can solve the problems of low conversion rate and poor repeatability of electrotransformation, and achieve the effect of overcoming poor repeatability
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Embodiment 1
[0024] The invention discloses a method for electrotransformation of Lactobacillus casei. The method comprises cultivation of Lactobacillus casei, electrotransformation of Lactobacillus casei and recultivation after transformation, and the specific steps are:
[0025] Lactobacillus casei culture:
[0026] A. Take out a Lactobacillus casei strain preserved by adding glycerol from the low-temperature freezer, streak it on the MRS solid medium, and invert overnight at 37°C anaerobically;
[0027] B. Use a sterile inoculation loop to pick the colony grown on the MRS into 3-5mL MRS liquid medium, and culture it overnight at 37°C;
[0028] C. On the second day, transfer 1 mL of the bacterial liquid grown the previous day to 50 mL of MRS liquid, grow statically at 37°C until the ultraviolet absorption value at 600nm of the bacterial liquid is 0.5-0.6, and prepare Lactobacillus casei for electroporation;
[0029] Electroporation of Lactobacillus casei:
[0030] D. Transfer Lactobaci...
Embodiment 2
[0041] Experiment purpose and method: In order to compare the changes in the conversion rate of Lactobacillus before and after the optimization of the experimental method, the operation method after optimization is as described in Example 1, and the operation method before optimization is as follows:
[0042] Insert a single colony of Lactobacillus into the MRS culture medium, culture it overnight at 37°C, then insert 2% of the inoculum into 20ml of the MRS culture medium, culture it at 37°C for about 4 hours, and reach the UV absorption value at 600nm of the culture medium 0.15-0.18, centrifuge to collect the cells, and wash the cells twice with an equal volume of ice-cold shock buffer. Cultures were suspended in 200 μl of shock buffer.
[0043] Take 1 μl of plasmid DNA and mix it with 40 μl of ice-cold cell suspension, transfer it into the electric shock cup and put it on ice for 5 minutes, set the voltage to 1175kV, the resistance to 100Ω, and the capacitance to 25μF, and t...
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