Difference between revisions of "Part:BBa K2717006:Experience"
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According to the western blot analysis, the glucose dehydrogenase has been well expressed (Fig. 1a). We inoculated the successfully expressed strains in the anti-antibiotic medium, induced by IPTG, and periodically measured the OD600 value of the bacterial liquid to draw a growth curve(Fig.1b and 1c)The results confirmed that gdh increased the environmental capacity of the cells and promoted the growth of the cells. The specific experiments and data processing can be found in our wiki. | According to the western blot analysis, the glucose dehydrogenase has been well expressed (Fig. 1a). We inoculated the successfully expressed strains in the anti-antibiotic medium, induced by IPTG, and periodically measured the OD600 value of the bacterial liquid to draw a growth curve(Fig.1b and 1c)The results confirmed that gdh increased the environmental capacity of the cells and promoted the growth of the cells. The specific experiments and data processing can be found in our wiki. | ||
| + | https://static.igem.org/mediawiki/2018/b/b6/T--BNU-China--_R1.png | ||
| − | + | Fig.14 Overexpression of glucose dehydrogenase | |
| − | + | A). Design of the experimental plasmid. | |
| − | Fig. | + | B) Western-blot image of the overexpressed GDH, from left to right are: bacteria with the recombinant plasmid, bacteria with the wild-type plasmid and wild-type bacteria (no plasmid transformed). |
| + | C) the growth curve with a dilution ratio of 1:10, y-axis represents OD600 and x-axis represents time. Through all the experiments, M9 media is used. The original OD600 is 1.60. | ||
| + | D) The bacteria growth curve after a dilution of 1:50, original OD600 is 0.58. | ||
Latest revision as of 21:27, 17 October 2018
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Applications of BBa_K2717006
According to the western blot analysis, the glucose dehydrogenase has been well expressed (Fig. 1a). We inoculated the successfully expressed strains in the anti-antibiotic medium, induced by IPTG, and periodically measured the OD600 value of the bacterial liquid to draw a growth curve(Fig.1b and 1c)The results confirmed that gdh increased the environmental capacity of the cells and promoted the growth of the cells. The specific experiments and data processing can be found in our wiki.
Fig.14 Overexpression of glucose dehydrogenase
A). Design of the experimental plasmid.
B) Western-blot image of the overexpressed GDH, from left to right are: bacteria with the recombinant plasmid, bacteria with the wild-type plasmid and wild-type bacteria (no plasmid transformed).
C) the growth curve with a dilution ratio of 1:10, y-axis represents OD600 and x-axis represents time. Through all the experiments, M9 media is used. The original OD600 is 1.60.
D) The bacteria growth curve after a dilution of 1:50, original OD600 is 0.58.
According to the method obtained in the literature, we determined the plasmid copy number in the bacteria at different time to verify the plasmid retention function of gdh. The experimental results confirmed that gdh has a small plasmid retention ability, but it did not achieve the expected effect, so we designed BBa_K2717010, BBa_K2717011 and BBa_K2717012 further detect the plasmid-retention capacity of gdh. The specific experiments and data processing can be found in our wiki.
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