Enginnering
We constructed three plasmids this year and we proved they could all work successfully. The aim we constructed these three plasmids is to use E.coli BL21(DE3) as a tiny factory to produce sericin and chitosan. So we considered to put the gene in a pET28a(+) plasmid, which is specifically used for E. coli expression. The construction result is proved by DNA sequencing. Then we changed the IPTG concentration, temperature and cultivating time to find the most proper condition. Because the protein we produced are linked with His-tag, so we use his-antibody to perform western blot to confirm its expression.
Part:BBa_K3874000
pET-28a(+)-Ser-01
Western Blot Data prove IPTG 0.5mM in 18℃ and a cultivation time of 24 hours is the most proper conditions here, so we used this set of conditions to conduct our experiment.
Part:BBa_K3874001
pET-28a(+)-Ser-08
Western Blot Data prove IPTG 0.5mM in 18℃ and a cultivation time of 20 hours is the most proper condition here, so we use this condition in latter experiment.
Part:BBa_K3874002
pET-28a(+)-Chitosan
Western Blot Data prove IPTG 0.5mM in 37℃ and a cultivation time of 4 hours is the most proper condition here, so we use this condition in latter experiment.
