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Fig. 1 | Biotechnology for Biofuels

Fig. 1

From: Identification of two integration sites in favor of transgene expression in Trichoderma reesei

Fig. 1

Expression of 2A self-cleavage peptide linked poly-protein gene in T. reesei. a Schematic diagram of the expression cassette of 2A peptide linked lipase gene and DsRed gene. Pcbh1 represents cbh1 promoter; Tcbh1 indicates the cbh1 terminator; sp is the abbreviation of signal peptide; His indicates 6×His tag; 2A indicates 2A peptide; LipA-F and Red-R are primers for genotyping the integration. b Representative image showing intracellular expression of RFP which was used to indicate the expression level of extracellular lipase. Images were taken under 40× objective lens using fluorescent microscope-Leica DMI4000B equipped with a mercury lamp. The excitation and emission spectra were 557 and 585 nm, respectively. c Lipase activities of three independent transformants and Tu6 strains in the supernatant of 96 h post-inoculation in MM media with 2% of lactose (w/v) and 5 mM uridine. The lipase activity was normalized to IU/g biomass. All the values in the figure are the mean value of three replicates. Error bars are the standard deviation (SD) between these replicates. d SDS-PAGE analysis of the supernatant from the same culture as in c. e Western blot analysis of expressing His-tagged lipA-2A-DsRed fusion protein in T. reesei. Samples were as same as that in d

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