Difference between revisions of "Part:BBa K4621150"

Revision as of 16:11, 5 October 2023

It consists of the chi18A hypothetical promoter and EctOperon

Usage and Biology

EctOperon contains ectA, ectB, ectC and ectD and can produce 5-Hydroxyectoine from lysine.

chi18A hypothetical promoter is a whole functional sequence including the promoter of chitinase Chi18A gene, which was cloned from SCUT-3 gene. In our project, through transcriptome analysis, qPCR and other means, we found that the gene expression of chitinase Chi18A was up-regulated when SCUT-3 was in chitin medium, and we speculated that its promoter chi18A hypothetical promoter may have properties that can be induced by chitin.

Testing and validation

We ligated the chi18A hypothetical promoter with EctOperon and overexpressed it in Streptomyces SCUT-3 using an integrated plasmid. The modified strain was inoculated in 40 mL LB medium and fermented at 40℃ and 220 rpm for 120h. The production of Ectoine and 5-Hydroxyectoin in the presence or without chitin (the first day of fermentation) was detected to evaluate the chitin inducability of the promoter. The results are shown in FIG. 1.

Fig.1 Fermentation of products with or without the addition of an inducer（chitin) (a).Ectoine (b).5-Hydroxyectoin(72h)

Sequence and Features