Grant: $156,083 - National Institutes of Health - Jul. 17, 2009
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Award Description: The proposed program will provide a genuine, goal-oriented research experience for high school students and teachers, introduce students to a potential career path in biomedical science, extablish relationships with local high school science teachers that could be the foundation of a long-term program, and finally accomplish an important research objective relevant to embryogenesis, developmental defects and diabetes.
Project Description: The goals of the work supported by this Supplement is to acquire high-resolution crystal structures of novel DNA-binding transcrlption factors of the panceras. Progress: To derive protein crystals of the trimeric complex, the students and teachers subcloned the cDNAs for the three individual subunIts (PTF1A, RBPJ and HEB, plus the isoform RBPJL) of the complex into a state-of-the-art expression vector for bacterial hosts. In addition several truncated variants of the subunits were also cloned (see deliverables). The vectors were transformed into TOP10 competent E. coli and identified by blue/white colony selection. The progress of cDNA cloning was monitored by Restriction endonuclease digestion and analytic gel electrophoresis. The final, correct expression constructs were sequenced across the regions modified by the subcloning, including the cDNA, and crucial elements of the vector including the bacterial promoter. These cDNA expression are now available for transformation into E. coli strain BL21 pLysS for high level production of protein. In forthcoming experiments, the expressed proteins will be purified and used to establish conditions for crystallization. Deliverables: A total of 29 cDNA expression plasmids for the following full length transcription factor subunits (1) PTF1A, (2) RBPJ, (3) RBPJL, (4) E12 and the following crystallization variants: (5-21) PT1A-nested truncations, (22-23) RBPJ-truncations, (24-25) RBPJL-truncations, and (26-29) E12 nested truncations.
Jobs Summary: N/A (Total jobs reported: 0)
Project Status: Less Than 50% Completed
This award's data was last updated on Jul. 17, 2009. Help expand these official descriptions using the wiki below.