Difference between revisions of "Part:BBa K3771098"

Revision as of 03:57, 18 October 2021

PsoxS-Riboj-RCP-J23101-SoxR​

Description

This composite part consists of promoter P_soxS Mutant 3 (BBa_K2862010), genetic insulator RiboJ (BBa_K1679038), RCP (BBa_K592012), constitutive promoter J23101 (BBa_J23101), and the soxR gene (BBa_K223000).

The soxR and the soxS gene are components of the soxRS regulon, which is important for E. coli to sense and respond to the oxidants. When SoxR protein is fully oxidized, it becomes a powerful transcription activator of the soxS promoter (P_soxS) (up to 100-fold), leading to the expression of the downstream gene^[1]. RiboJ is a genetic insulator that can increase the expression of insulated genes. RCP serves as a reporter to help us pick up the correct colony.

Usage and Biology

This composite part was ligated with the pSAA vector and transformed into E. coli. We conducted colony PCR to verify whether E. coli uptake the correct plasmid. The size of the PCR product was as expected.

Fig.1. The electrophoresis result of colony PCR. M: Marker; Lane 1: pSAA-P_soxS-sfgfp (990 bp); Lane 2, 3: pSAA-P_soxS-RiboJ-rcp-J23101-soxR (1103 bp).​

References

Pomposiello PJ, Demple B. Redox-operated genetic switches: the SoxR and OxyR transcription factors. Trends Biotechnol. 2001;19(3):109-114. doi:10.1016/s0167-7799(00)01542-0

Sequence and Features