Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jianyi Huang
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-17 08:39:48
pT7+pTet+RBS+tetX-GFP(fusion protein)+DT
| Types | DnaRegion |
| Roles | engineered_region Reporter |
| Sequences | BBa_K2150019_sequence (Version 1) |
Description
Two promoters are used, a constitutive promoter pTet and a T7RNAP specific promoter pT7. The fusion protein is tetX and GFP with a 3*GGGGS linker so that the expression level of tetX can be accurately reported by the fluorescence intensity of GFP. By testing the fluorescence intensity, we found that in the absence of T7 promoter, these two promoters exhibit higher efficiency than single pTet; the fluorescence intensity is lower that BBa_K2150018, in which the fusion protein is substituted by single tetX. With pT7, expression of GFP can be amplified while with pTet, expression of GFP can be repressed by tetR.Notes
Fusion protein is designed to report the accurate expression level of tetX. With pT7 promoter, expression of tetX-GFP can be amplified.Source
pT7 PCR amplified from BBa_K525998,pTet from BBa_R0040,RBS from BBa_B0034,DT from BBa_B0015As for fusion protein tetX-GFP, tetX is the same as (xxx), the protein linker is provided by our advisor Li Hua, and the GFP is from BBa_E0040. We combine this three parts using overlap extension polymerase chain reaction (OE-PCR).
| Access | Instance | Definition |
| public public public public public | BBa_K2150031 BBa_R0040 BBa_B0034 BBa_K2150013 BBa_B0015 | pT7 p(tetR) BBa_B0034 tetX-GFP BBa_B0015 |