Part:BBa_K1841000:Design
nisin resistancy
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Our goal is design a composite gene that can constant produce nisI by E. coli and L. casei.
The promoter pUO19 contained - 10 and -35 regions similar to the consensus sequences of E. coli and Lactobacillus promoters. This provide us a convenient way to test the function of the nisI in the E. coli rather than experiment on the L. casei we can not familiar with. As the same reason we use the RBS from L. casei, but it still function well in E. coli to achieve our goal. The most important gene of this part nisI. Nisin-producing Lactococcus lactisstrains show a high degree of resistance to the action of nisin, which is based upon expression of the self-protection (immunity) genes nisI, nisF, nisE, and nisG. NisI acts as a nisin-sequestering protein and that NisFEG acts as a nisin exporter that expels nisin molecules from the cytoplasmic membrane into the environment.
It means we can use a proper way to make sure the transformation success rate instead of using antibiotic which is banned in the food industry.
Source
total synthesis by IDT
References
1. Torsten, S., Stefan, H., Irina, S., and K. D. Entian. Function of Lactococcus lactis Nisin Immunity Genes nisI and nisFEG after Coordinated Expression in the Surrogate Host Bacillus subtilis. The Jurnal of Biological Chemistry, USA. , Vol. 278, pp. 89 -94 (2003) 2. Pilar Garcia, Victoria Bascaran, Ana Rodriguez, and Juan Evaristo Suarez. Isolation and characterization of promoters from the Lactobacillus casei temperate bacteriophage A2 (1997) 3. SUNGMIN F. KIM, SEUNG J. BAEK, AND M. Y. PACK. Cloning and Nucleotide Sequence of the Lactobacillus casei Lactate Dehydrogenase Gene (1991)