《Table 2 Strains used in this studya)》

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《Rational design for fungal laccase production in the model host Aspergillus nidulans》

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a) TXX，original transformant.

To construct the overexpression plasmid，the tryptophan synthase（TrpC）terminator sequence was amplified from the genomic DNA of A.nidulans RDIT9.32 using the primers TrpC-for and-rev and integrated into plasmid pJMP9.1 to create pYWL14.1 with the gpdA promoter and trpC terminator using the quick-change strategy（Bok and Keller，2012）.The pslcc gene was amplified from the genomic DNA of P.sanguineus mk528 using the primers Ps528 lcc-for and-rev and integrated into plasmid pJMP14.1 to create pYWL15 using the abovementioned protocol.The laccase expression cassette，including its own promoter，pslcc gene and its own terminator，was amplified from the genomic DNA of P.sanguineus mk528 using the primers Ps528PLT-F-Not I/-R-Avr II or Ps528PLT-for/-rev.The former pslcc expression cassette was digested with the restriction enzymes Not I and Avr II，then the Not I-Avr II fragment of the cassette was cloned into the Not I and Avr II sites of pRG-AMA1-Not I to create pYWL16.The latter was then integrated into plasmid pYWL14.1 to create pYWL17.1 using the quick-change strategy.To construct the overexpression plasmid with different marker gene cassettes，the plasmid pWL93.4 was constructed by integrating the trpC terminator into pWY25.16.Then，plasmids pWL94.8 and pWL95.6were constructed by replacing the marker gene cassette AfriboB（A.fumigatus riboflavin biosynthesis gene）and pyroA based on pWL93.4.Finally，the overexpression plasmids pWL97.2，pWL99.2 and pWL100 of laccase with the gpdA promoter and different marker genes were constructed based on pYWL95.6，pYWL94.8 and pYWL93.4，respectively.To construct the overexpression plasmid with the aflR promoter，the aflR promoter sequence was amplified from the genomic DNA of A.nidulans RJMP1.59 using the primers aflR-promoter F and R and integrated into plasmid pYWL17.1 to create pYWL53.1 with the laccase overexpression cassette，including the aflR promoter，pslcc gene and trpC terminator using the quick-change strategy.We amplified the aflR promoter and pslcc gene cassette and integrated it into plasmid pYWL94.8 to create pYWL55.3. This construct was confirmed by PCR with primers and then sequenced.Plasmids were isolated by using the Plasmid Mini Kit I（OMEGA，USA）.The oligonucleotide sequences for the PCR primers are given in Table S2 in Supporting Information.