Cotton bollworm single particle embedded nuclear polyhedrosis virus shuttle expression carrier and preparation method
A technology of shuttle expression vector and karyotype polyhedron, applied in the fields of molecular biology and bioengineering, can solve the problems of low yield of recombinant virus, time-consuming, complicated operation of plaque purification method, etc.
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[0015] The steps are as follows:
[0016] 1. Activation of DH10B recipient strain 1 containing HZ8 (HaBacmid) and helper plasmid. Inoculate DH10B recipient bacteria stored at -80°C on LB solid medium containing kanamycin (50 μg / ml) and tetracycline (50 μg / ml), culture at 37°C for 12-16 hours, pick a single colony and inoculate into On 3 mL of LB liquid medium containing kanamycin (50 μg / ml) and tetracycline (50 μg / ml), culture at 37° C. for 24 hours, and prepare competent recipient bacteria according to conventional methods.
[0017] 2. Activation of DH5α bacteria containing HapFastPhP10 plasmid and extraction of HapFastPhP10 plasmid 2. Inoculate DH5α recipient bacteria stored at -80°C on LB solid medium containing gentamicin (50 μg / ml), culture at 37°C for 12-16 hours, pick a single colony and inoculate into 3 mL containing gentamicin ( 50 μg / ml) LB liquid medium, cultured at 37° C. for 16 hours, and extracted the HapFastPhP10 plasmid according to the conventional method fo...
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