Team:BUCT-China/Contribution
Contribution
The new part
We registered 4 parts to share our results and hope to be used by other igemers.
BBa_K3197001 is a promoter .
BBa_K3197002 is a RBS.
BBa_K3197003 encodes a kind of oxidase .This oxidase could degrade polyolefin into shorter carbon chain, which is of great benefit to the management of polyolefin plastic pollution and also of great significance to the degradation of other polymers.
BBa_K3197004 is a terminator .
They are all got from Microbulbifer hydrolyticus, by sequencing.
The characterization of exiting part BBa_J04450
We characterized the function of a part BBa_J04450 . We used pSB1C3 plasmids to introduce it into Trans 10 for culture, observe its growth, and plot the growth curve.
We contribute to the characterization of part BBa_J04450 by introducing it in Trans10 to observe its growth and draw its growth curve.
Experimental Design
1.The target gene was obtained by PCR cloning.
2. Digest the gene and plasmid pSB1C3 with enzymes Dpn,IEcoR,Pst I.
3.Ligate the digested gene fragment and plasmid pSB1C3 with T4 enzyme.
4.Transfer the ligation product to Trans 10 by thermal conversion.
5.Plate the transformed strain and culture at 37℃ for 12h.
6.Pick a single colony from the plate to make a bacterial solution. The cells were cultured separately in 24 tubes, and every tube was measured the absorbance in order for each 1 hour.
Results
The growth of the plate colonies was observed as shown in the Figure a. We could see that an appropriate amount of colonies grew on the plate, indicating that the target gene fragment was successfully cloned into Trans 10.
Figure a： 12 hours old transformed Trans 10 strains.
We quantitatively described the growth curve of Trans10 single colony in medium. (Figure b) According to the growth rate constant of the bacteria, we could divide 24h into three different growth phases：0~2h is lag phase; 2~ 15h is an exponential phase; 15~ 24h is a stationary phase.
Figure b：The growth curve of a single colony from the plate.