Sterility:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Hood:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Electroporation:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Article Title: Context-Dependent Modulation of Replication Activity of Saccharomyces cerevisiae Autonomously Replicating Sequences by Transcription Factors
Article Snippet: .. The competent yeast cells were mixed with 1 mg each of the control plasmid DNA (pHSG3982), reporter plasmid DNA, and effector plasmid, transferred into an ice-cold disposable electroporation cuvette (0.2-cm gap) and pulsed at 1.5 kV, 25 mF, 400 V with a GENE Pulser (Bio-Rad). ..
Article Title: On the Evolutionary Origin of Cyclooxygenase (COX) Isozymes
Article Snippet: .. 250 l of cell suspension (1 107 cells/ml) was mixed with 400 ng of plasmid DNA and 50 g of salmon sperm DNA in a disposable electroporation cuvette and was subjected to an electric discharge of 180 V using a Bio-Rad Gene Pulser at 970-microfarad capacity. .. For activity assay, transfected cells were grown at 28 °C for 72 h. Tunicamycin treatments of COS-7 cells transfected with pCGCOX-A, pCG-COX-B, or pCG-RabCOX-2 constructs were performed by adding tunicamycin (Sigma) to the media at final concentration of 0.3 or 1.0 g/ml 4 h post-transfection.
Plasmid Preparation:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Article Title: Context-Dependent Modulation of Replication Activity of Saccharomyces cerevisiae Autonomously Replicating Sequences by Transcription Factors
Article Snippet: .. The competent yeast cells were mixed with 1 mg each of the control plasmid DNA (pHSG3982), reporter plasmid DNA, and effector plasmid, transferred into an ice-cold disposable electroporation cuvette (0.2-cm gap) and pulsed at 1.5 kV, 25 mF, 400 V with a GENE Pulser (Bio-Rad). ..
Article Title: On the Evolutionary Origin of Cyclooxygenase (COX) Isozymes
Article Snippet: .. 250 l of cell suspension (1 107 cells/ml) was mixed with 400 ng of plasmid DNA and 50 g of salmon sperm DNA in a disposable electroporation cuvette and was subjected to an electric discharge of 180 V using a Bio-Rad Gene Pulser at 970-microfarad capacity. .. For activity assay, transfected cells were grown at 28 °C for 72 h. Tunicamycin treatments of COS-7 cells transfected with pCGCOX-A, pCG-COX-B, or pCG-RabCOX-2 constructs were performed by adding tunicamycin (Sigma) to the media at final concentration of 0.3 or 1.0 g/ml 4 h post-transfection.
Suspension:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Article Title: On the Evolutionary Origin of Cyclooxygenase (COX) Isozymes
Article Snippet: .. 250 l of cell suspension (1 107 cells/ml) was mixed with 400 ng of plasmid DNA and 50 g of salmon sperm DNA in a disposable electroporation cuvette and was subjected to an electric discharge of 180 V using a Bio-Rad Gene Pulser at 970-microfarad capacity. .. For activity assay, transfected cells were grown at 28 °C for 72 h. Tunicamycin treatments of COS-7 cells transfected with pCGCOX-A, pCG-COX-B, or pCG-RabCOX-2 constructs were performed by adding tunicamycin (Sigma) to the media at final concentration of 0.3 or 1.0 g/ml 4 h post-transfection.
Transfection:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Centrifugation:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Incubation:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Inverted Microscopy:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Expressing:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Enzyme-linked Immunosorbent Assay:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
High Performance Liquid Chromatography:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Recombinant:Article Title: Recombinant varicella-zoster virus (VZV) vaccine
Article Snippet: .. In a sterile laminar flow bench, a perforation voltage of a gene pulse generator Xcell (Bio-Rad) was set as follows: 300 V, 900 μF single pulse, and infinite resistance; a disposable electroporation cuvette (Bio-Rad) with a gap of 4 mm was taken out and added with 40 μg of a linearized plasmid DNA (100 μ1) and 0.7 ml of a CHO K1 cell suspension (1.5×107 cells/ml), and the linearized plasmid VZV gE-Fc-straight was directly transfected into CHO K1 cells by electrotransfection; the cells in the electroporation cuvette were transferred into a triangular culture flask, 30 ml of a CD CHO medium (GIBCO) was added, and the cells were cultivated in a shaker at 36° C. to 37° C., 5% CO2, and 135 rpm for 24 h; then the cells were collected by low-speed centrifugation and inoculated into a 50 μM MSX-containing CD CHO medium (without glutamine) instead; a resulting cell suspension was transferred into a 96-well flat-bottom culture plate by limiting dilution, and the culture plate was incubated in a 37° C. and 10% CO2 incubator; the cells were observed under an inverted microscope, and monoclonal cell wells were marked; then the monoclonal lines with high expression were screened out by ELISA (goat anti-human IgG+expression product VZV gE-Fc+goat anti-human IgG-HRP) and protein A HPLC; the screened lines were continuously subcultivated and tested, and finally 3 cell clone lines with high expression of the target gene were obtained, with clone numbers of 5B3, 8D8, and 12C3; and an expression level of the recombinant protein in the culture supernatant was detected according to the feed test and HPLC, and the clone line 5B3 was selected for scale-up experiment. ..
Control:Article Title: Context-Dependent Modulation of Replication Activity of Saccharomyces cerevisiae Autonomously Replicating Sequences by Transcription Factors
Article Snippet: .. The competent yeast cells were mixed with 1 mg each of the control plasmid DNA (pHSG3982), reporter plasmid DNA, and effector plasmid, transferred into an ice-cold disposable electroporation cuvette (0.2-cm gap) and pulsed at 1.5 kV, 25 mF, 400 V with a GENE Pulser (Bio-Rad). ..
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