Overview
Our team aims to mitigate soil salinization with microorganisms, specifically Clostridium tyrobutyricum. By genetically engineering C. tyrobutyricum, we inserted a pMTL82151 plasmid containing DNA segments encoding the dps and aceE proteins, thus reconstructing the main metabolic pathway of C. tyrobutyricum and allowing it to survive and reproduce in aerobic environments. Genetically engineered C. tyrobutyricum produces butyric acid, which can be used to neutralize salt-affected soils, as well as replacing antibiotics, for it can promote the bowel health of domestic animals.
We have reached the following achievements.
1.Characterization of the part BBa_K1033933
We obtained the plasmid vector containing this part, pET-29a-aspink plasmid, from iGEM team iGEM13_Uppsala. We transformed this plasmid into E.coli strain BL21, and successfully induced aspink expression. We extracted and purified the aspink protein by sonication, ultrafiltration, and Ni-affinity chromatography. With bioinformation analysis using Swiss Model, we predicted the structure of aspink protein and analyzed detailed characteristics of the protein. The relative molecular mass was measured by mass spectrometry.
2.Plasmids expressing Dps and aceE protein
We constructed pMTL-Pthl-Dps, pMTL-Plac-Dps, pMTL-Pthl-aceE and pMTL-Plac-aceE to express Dps and aceE proteins with Pthl and Plac promotors, respectively. These plasmids may be of help in engineering other strict anaerobes to facultative anaerobes. Future iGEM teams that need to use these genes to realize genetic engineering of strict anaerobes may utilize these plasmids.
