Part:BBa_K2224001:Experience
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Applications of BBa_K2224001
Part Description:
This is the experiment report of SMS_Shenzhen team in August, 2017 BBa_K2224001 is a composite part with the backbone pSB1C3. The CDS of this part is a chitinase originate from verticillium lecanii, a kind of entomogenous fungus. With a GeneBank identifier DQ412944 in NCBI. This enzyme helps catalyze chitin. SMS students optimized the codons and removed introns to fit the original sequence into E. coli expression system.
Experiment Abstract
The original sequence DQ412944 was published on NCPI. SMS students have processed the CDS. Designed primer with restriction sites, modified the CDS for standardization. We then synthesized this plasmid with the backbone pSB1C3.
We used a Chitinase activity measurement kit to measure the activity of this chitinase quantitatively. The activity was defined by the amount of N- acetyl glucosamine that produced by Chitinase' s hydrolysis to chitin divided by the amount of cells. Since the product, N- acetyl glucosamine, can lead to light absorbtion peak at 585nm and the light absorption can be detected by Spectrophotometer, we can figure out the relative activity of enzyme DQ412944. Enzyme activity in TOP10 E. coli expression system related to time shown below. (ABS is for absorbance, U is for active unit)
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