Team:BJ101ID/Result.html

What we have made Part1

We use E.coli BL21(DE3) as a tiny factory to express sericin and chitosan. According to our design, we constructed three plasmids this year and we proved they could all work successfully.

Part:BBa_K3874000 pET-28a(+)-Ser-01

Part:BBa_K3874001 pET-28a(+)-Ser-08

Part:BBa_K3874002 pET-28a(+)-Chitosan

We considered to put the gene in a pET28a(+) plasmid, because it 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 cultivating for 24h is the most proper condition and we use this condition in latter experiment.

Part:BBa_K3874001 pET-28a(+)-Ser-08

Western Blot Data prove IPTG 0.5mM in 18℃ and cultivating for 20h is the most proper condition and we use this condition in latter experiment.

Part:BBa_K3874002 pET-28a(+)-Chitosan

Western Blot Data prove IPTG 0.5mM in 37℃ and cultivating for 4h is the most proper condition and we use this condition in latter experiment. After we find the most proper condition, we use it to cultivate E.coli in a larger volume and we get more outcome to do the second part of our experiment.