BME494 Project Group9
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ABSTRACT
BACKGROUND
The proposed design has the potential to be used as a simple and efficient diagnostic test.
PROOF OF CONCEPT DESIGNIn the presence of 25(OH)D3 in blood, the co-transcription factor and promoter for the CYP21B1 are activated. When the promoter is activated, it expresses the enzyme that causes a chemical change to to the active form of Vitamin D. This active form, 1-25-DiHydroxyVitamin D3, is created in the kidney. If the promoter is activated, then it will express the enzyme that chemically changes to the active form of vitamin D in the kidney.
The genes for the CYP27B1 co-transcription factors and promoter were provided via GenBank (http://www.ncbi.nlm.nih.gov/genbank/). Prior to primer creation, it was determined if there was the need for site directed mutagenesis. Fortunately, through the aid of Nebcutter, it was determined that the original gene set was ok to use as is. The forward primer: 5'-ctttctttcgcgagcacc-3' The reverse primer: 5'-tataaacgcagaaaggc-3' One pre-determined complication withthis primer design is the lack for GC clamps. In testing, this could present future problems during transformation and amplification. The pre-existing Biobrick part BBa_101015 is used and includes a RBS, GFP, and a terminator. The 2008 Minnesota iGem team has biobricked this part and found it successful in their projects, thus it is expected to work correctly in our system.
File:C:\Documents and Settings\Robbia\My Documents\My Pictures\Plasmid Assembly.JPG
FUTURE VISION: D-Vitameter: Ideally this would be a device that would benefit from the proposed technology. This compact and portable device would allow patients to get a reading of their ability to produce Vitamin D by just using one drop of blood. The basic components of this small device would be an enclosed box with a small polycarbonate window. The device will allow to introduce a sample in a microscope slide and shine a UV light to detect presence of 25(OH).
TESTING
When 25(OH)D3 is present, ideally there would be a 100% GFP concentration
HUMAN PRACTICES•The meter is practical because the blood sample is relatively small and can be extracted at any time. Normal tests for Vitamin D deficiency require laboratory procedures and time.
OUR TEAM
Senior Biomedical Engineering, SBHSE, I am interested in learning the general concepts of synthetic biology and its applications, I will be graduating this semester! http://openwetware.org/wiki/User:Carolina_Tostado
Senior in Biochemistry.
Junior in Biomedical Engineering, SBHSE.
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