Alkaline denaturation of pJQCtet, pJQBtet, pJQDtet, and pJQCBDtet was performed as described earlier [51], and the electroporation was carried out using a gene pulser II (BioRad, Harcules, CA).
The plasmid extracts were electroporated into E. coli DH5α by using a Gene Pulser II apparatus (Bio-Rad, Hercules, CA), and the transformants were selected on Luria-Bertani agar plates containing amikacin (30 μg/mL).
Sac I was used to linearize 5 10 μg pPIC9K-N-APP plasmid DNA, and linear DNA was then transformed into the competent P. pastoris GS115 cells using a Gene Pulser system (Bio-Rad; conditions used: 1.5 kV, 200 Ω, 25 μF, and 4.5 ms).
Transfection of pDEST26 or pDEST26-GSTP1 in control- and HPV-16 E7-infected MCF-7 cells was done by electroporation using the Gene Pulser II apparatus (Bio-Rad) according to Manufacturer's instructions.
The recombinant plasmid (pYMAb2:: blaOXA-72) and control plasmid (pYMAb2) were then transformed into the kanamycin-susceptible Ab290 strain by electroporation using an gene pulser electroporator (Bio-Rad, Hercules, CA) and 2 mm electrode gap cuvettes.
pPICXYN was linearized with Sal I and electroporated into P. pastoris using a Gene Pulser electroporator (Bio-Rad, http://www.bio-rad.com), following the manufacturers' instructions.
To establish SOCS1-overexpressing HACs, pShuttle2-SOCS1 or empty vector was electrotransfected by using a Gene Pulser Xcell System (Bio-Rad, Hercules, CA, USA) under the condition of 50-V and 2-ms pulse (4-mm cuvette).
One microgram of PCR product was added to 100 μl of BL21 (DE3) competent cells containing the lambda red recombinase expression plasmid pKD46, and electroporation was performed using a Gene Pulser II transfection apparatus (Bio-Rad Laboratories, HerCAles, CA, USA) at 25 μF, 2.4 kV and 250 Ω.
Electroporation was performed using a Gene Pulser Xcell system (Bio-Rad) at 0.85 kV twice with a 5 s interval.
