β-lactamases and their coding genes and their applications
A lactamase and gene technology, applied in the field of biotechnology, can solve problems such as carbapenem antibiotic pollution and carbapenem antibiotic metabolism, and achieve the effect of removing antibiotic pollution
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Embodiment 1
[0040] This example is used to illustrate the resistance of Escherichia coli transformed with the β-lactamase coding gene provided by the invention to β-lactam antibiotics
[0041] (1) Preparation of competent E.coli BL21 cells
[0042] 1) Streak culture competent Escherichia coli BL21 on LB solid medium, culture at 37°C for 12-16 hours.
[0043] 2) Pick fresh monoclonal colonies, inoculate them into 4ml LB liquid medium, culture with shaking at 37°C for 12-16 hours.
[0044] 3) On the third day, add 1ml of bacterial solution to 100ml of LB liquid medium at a ratio of 1:100, shake at 37°C and 220rpm for about 2 hours, and measure OD every half an hour 600 , when OD 600 When it reaches 0.5, the culture is stopped.
[0045] 4) After 30 minutes of ice bathing the bacterial solution, the bacterial solution was dispensed into 50 ml pre-cooled centrifuge tubes, and centrifuged at 4500 rpm for 10 minutes at 4°C.
[0046] 5) Discard the supernatant, add a small amount of pre-cooled ...
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