Team:NPU-CHINA/Contribution
CONTRIBUTIONS
In our team’s project this year, we use the pHT01 plasmid as vector. This plasmid contains a lactose operon suitable for Bacillus subtilis. The promoter of this lactose operon is Pgrac （BBa_K1628202）. We found some information related to this promoter when we searched for the information in the early stage, and noted it under the part page.
Information Adding：
NPU-CHINA 2021:Through the information searched by our team members[1], according to figure 3(A) pNDH33 and the DNA sequence of the Pgrac promoter (in capital letters) including the upstream AT-rich UP element, the lac operator (lacO; in capital letters) and the ribo-some-binding sequence (underlined) (showed in Figure 3)
NPU-CHINA 2021:According to the literature[2], we found Pgrac100 not only tightly controls the background expression level in E.coli , but also allowed high protein production levels at low IPTG concentrations(showed in Table 1).
Reference
[1]Phan TT, Nguyen HD, Schumann W. Novel plasmid-based expression vectors for intra- and extracellular production of recombinant proteins in Bacillus subtilis. Protein Expr Purif. 2006 Apr;46(2):189-95. doi: 10.1016/j.pep.2005.07.005. Epub 2005 Aug 9. PMID: 16125412.
[2]Phan, Trang & Linh Thuoc, Tran & Schumann, Wolfgang & Nguyen, Hoang. (2015). Development of Pgrac100-based expression vectors allowing high protein production levels in Bacillus subtilis and relatively low basal expression in Escherichia coli. Microbial cell factories. 14. 72. 10.1186/s12934-015-0255-z.
We encountered troubles in this year's project, especially the construction of recombinant plasmids and Bacillus subtilis transformation took us a lot of time, but finally found a comparatively stable solution. We organize our solutions into Protocol, and hope to help the team engaged in related research in the future.
Please see：Protocol and Lab work
The previous iGEM team in the northwest region jointly established a part base, which is operated and maintained by several teams in the northwest region. We used the free sequence synthesis service provided by Twist this year to synthesize several components and record them into Part Base.
Part_Base.pdf
CONTACT NPU CHINA
NPU-CHINA is the iGEM team of Northwestern Polytechnical University
Address: 127 West Youyi Road Beilin District, Xi'an,
Shaanxi 710072, China
Contact: npuigem@outlook.com
bilibili: NPU-iGEM
