pPICZαB

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	The PCR product lccC was digested with EcoRI and XbaI, ligated with the digested expression vector pPICZαB, and then transformed into the recombinant P. pastoris GS115 carrying pPICZαB competent cells. For production of laccase, the strain was transferred to YPD (yeast extract-peptone-dextrose) plates and incubated at 28oC for 3 days. BMGY (buffered glycerol complex) and YPD media were prepared according to the manual of the Easy Select Pichia Expression Kit (Invitrogen).

Protocol tips

The PCR product lccC was digested with EcoRI and XbaI, ligated with the digested expression vector pPICZαB, and then transformed into the recombinant P. pastoris GS115 carrying pPICZαB competent cells. For production of laccase, the strain was transferred to YPD (yeast extract-peptone-dextrose) plates and incubated at 28oC for 3 days. BMGY (buffered glycerol complex) and YPD media were prepared according to the manual of the Easy Select Pichia Expression Kit (Invitrogen).

Publication protocol

"The recombinant strain P. pastoris GS115-lccC was constructed and preserved in the Microbial Technology Research Laboratory, Nanjing Forestry University. While P. pastoris GS115-lccC was constructed, PCR primers for cDNA amplification of lccC were designed based
on the sequence of lac4 (GenBank Accession No. 212734). The PCR product lccC was digested with EcoRI and XbaI, ligated with the digested expression vector pPICZαB, and then transformed into the recombinant P. pastoris GS115 carrying pPICZαB competent cells.
For production of laccase, the strain was transferred to YPD (yeast extract-peptone-dextrose) plates and incubated at 28oC for 3 days. BMGY (buffered glycerol complex) and YPD media were prepared according to the manual of the Easy Select Pichia Expression Kit (Invitrogen)."

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