Manipulating Expression of LP502, PcyA, and Ho1 with pET28b and PETDuet Vectors in Chemically Competent Escherichia coli
Session Number
A13
Advisor(s)
Don Dosch, Illinois Mathematics and Science Academy C. Robyn Fischer, Illinois Mathematics and Science Academy
Location
A-113
Start Date
28-4-2016 9:15 AM
End Date
28-4-2016 9:40 AM
Abstract
When combined, the proteins Ho1, PcyA, and LP502 make up the light- harvesting complex, or phycobilisome (PCB), of cyanobacteria. These complexes are primarily responsible for the light absorption that takes place in photosystem II of photosynthesis. Expression vectors pET28b- and pETDuet code specifically for the LP502 protein and the Ho1 and PcyA proteins. By inserting the photosystem into Escherichia coli, we limit the number of outside factors that affect the light absorption. The goal of this study is to transform E. coli with vectors pETDuet and pET28b- in order to generate the three aforementioned proteins. These plasmids can be transformed individually into E. coli, but when attempting to insert them both into the bacteria at the same time, the transformation efficiency decreases drastically. The plasmids we used in this investigation were clones sent from our collaboration partners at RDFZ. By manipulating how the bacteria is transformed we are able to observe how the expression of the PCB is affected. Engineering the expression of these proteins in E. coli connects to research in the advancement of production efficiency and effectiveness of various biomaterials, especially biofuel
Manipulating Expression of LP502, PcyA, and Ho1 with pET28b and PETDuet Vectors in Chemically Competent Escherichia coli
A-113
When combined, the proteins Ho1, PcyA, and LP502 make up the light- harvesting complex, or phycobilisome (PCB), of cyanobacteria. These complexes are primarily responsible for the light absorption that takes place in photosystem II of photosynthesis. Expression vectors pET28b- and pETDuet code specifically for the LP502 protein and the Ho1 and PcyA proteins. By inserting the photosystem into Escherichia coli, we limit the number of outside factors that affect the light absorption. The goal of this study is to transform E. coli with vectors pETDuet and pET28b- in order to generate the three aforementioned proteins. These plasmids can be transformed individually into E. coli, but when attempting to insert them both into the bacteria at the same time, the transformation efficiency decreases drastically. The plasmids we used in this investigation were clones sent from our collaboration partners at RDFZ. By manipulating how the bacteria is transformed we are able to observe how the expression of the PCB is affected. Engineering the expression of these proteins in E. coli connects to research in the advancement of production efficiency and effectiveness of various biomaterials, especially biofuel